Flow cytometric analysis of the in situ accessibility of Escherichia coli 16S rRNA for fluorescently labeled oligonucleotide probes.
AuthorsFuchs BM,Wallner G,Beisker W,Schwippl I,Ludwig W,Amann R
JournalApplied and environmental microbiology
PubMed ID9835591
In situ identification of whole fixed bacterial cells by hybridization with fluorescently labeled, rRNA-targeted oligonucleotide probes is often limited by low signal intensities. In addition to an impermeability of the cell periphery and a low cellular rRNA content, the three-dimensional structure of the ribosome may hinder the access of oligonucleotides ... More
Mechanism of peptide-induced mast cell degranulation. Translocation and patch-clamp studies.
AuthorsLorenz D, Wiesner B, Zipper J, Winkler A, Krause E, Beyermann M, Lindau M, Bienert M
JournalJ Gen Physiol
PubMed ID9806967
Substance P and other polycationic peptides are thought to stimulate mast cell degranulation via direct activation of G proteins. We investigated the ability of extracellularly applied substance P to translocate into mast cells and the ability of intracellularly applied substance P to stimulate degranulation. In addition, we studied by reverse ... More
Quantitative polymerase chain reaction-based homogeneous assay with fluorogenic probes to measure c-erbB-2 oncogene amplification.
We describe a PCR-based assay for determining c-erbB-2 oncogene amplification in breast cancer in which we use the TaqMan system. Two fluorogenic probes anneal to the target between primers for c-erbB-2 and beta-globin genes and contain both a reporter dye (6-carboxy-fluorescein) and a quencher dye (6-carboxy-tetramethyl-rhodamine). During the extension phase ... More
DNA sequencing with dye-labeled terminators and T7 DNA polymerase: effect of dyes and dNTPs on incorporation of dye-terminators and probability analysis of termination fragments.
The incorporation of fluorescently labeled dideoxynucleotides by T7 DNA polymerase is optimized by the use of Mn2+, fluorescein analogs and four 2'-deoxyribonucleoside 5'-O-(1-thiotriphosphates) (dNTP alpha S's). The one-tube extension protocol was tested on single-stranded templates, as well as PCR fragments which were made single-stranded by digestion with T7 gene 6 ... More
Fluorescence polarization immunoassay IV. Determination of phenytoin and phenobarbital in human serum and plasma.
AuthorsLu-Steffes M, Pittluck GW, Jolley ME, Panas HN, Olive DL, Wang CH, Nystrom DD, Keegan CL, Davis TP, Stroupe SD
JournalClin Chem
PubMed ID6751601
Fluorescence polarization immunoassays for phenytoin and phenobarbital in human serum or plasma are described and shown to be clinically useful. No sample pretreatment, extraction, or phase separation is required. A determination can be made with less than 20 microL of sample in 15 min, with incubation at ambient temperature. Abnormal ... More
A novel fluorescent toxin to detect and investigate Kv1.3 channel up-regulation in chronically activated T lymphocytes.
AuthorsBeeton C, Wulff H, Singh S, Botsko S, Crossley G, Gutman GA, Cahalan MD, Pennington M, Chandy KG
JournalJ Biol Chem
PubMed ID12511563
T lymphocytes with unusually high expression of the voltage-gated Kv1.3 channel (Kv1.3(high) cells) have been implicated in the pathogenesis of experimental autoimmune encephalomyelitis, an animal model for multiple sclerosis. We have developed a fluoresceinated analog of ShK (ShK-F6CA), the most potent known inhibitor of Kv1.3, for detection of Kv1.3(high) cells ... More
Catalyzed reporter deposition, a novel method of signal amplification. II. Application to membrane immunoassays.
AuthorsBobrow MN, Shaughnessy KJ, Litt GJ
JournalJ Immunol Methods
PubMed ID1849153
'In a previous publication (Bobrow et al., J. Immunol. Methods (1989) 279-285), we described a novel signal amplification method, catalyzed reporter deposition (CARD), and its application to microplate immunoassays. The method utilizes the analyte-dependent reporter enzyme (ADRE) to catalyze the deposition of additional reporter onto the surface of a solid-phase ... More
Activated human T lymphocytes express a functional C3a receptor.
AuthorsWerfel T, Kirchhoff K, Wittmann M, Begemann G, Kapp A, Heidenreich F, Götze O, Zwirner J
JournalJ Immunol
PubMed ID11086104
'The C3a molecule is an anaphylatoxin of the C system with a wide spectrum of proinflammatory effects predominantly on cells of myeloid origin. In this study we investigated the expression of the high affinity receptor for C3a (C3aR) in human T lymphocytes using receptor-specific mAb. C3aR expression was detected in ... More
A fluorescence-based method for determining the surface coverage and hybridization efficiency of thiol-capped oligonucleotides bound to gold thin films and nanoparticles.
AuthorsDemers LM, Mirkin CA, Mucic RC, Reynolds RA, Letsinger RL, Elghanian R, Viswanadham G
JournalAnal Chem
PubMed ID11101228
'Using a fluorescence-based method, we have determined the number of thiol-derivatized single-stranded oligonucleotides bound to gold nanoparticles and their extent of hybridization with complementary oligonucleotides in solution. Oligonucleotide surface coverages of hexanethiol 12-mer oligonucleotides on gold nanoparticles (34 +/- 1 pmol/cm2) were significantly higher than on planar gold thin films ... More
pH- and ionic strength-dependent fusion of phospholipid vesicles induced by pardaxin analogues or by mixtures of charge-reversed peptides.
AuthorsRapaport D, Hague GR, Pouny Y, Shai Y
JournalBiochemistry
PubMed ID8461295
'The fusogenic properties of the neurotoxin paradaxin and eight of its analogues with small unilamellar vesicles (SUV), composed of egg phosphatidylcholine and phosphatidylserine (PC/PS), were investigated. Fusion was demonstrated by a lipid-mixing assay and by an increase in vesicle size as revealed by electron microscopy. The lipid-mixing assay was performed ... More
Aggregation and organization of pardaxin in phospholipid membranes. A fluorescence energy transfer study.
AuthorsRapaport D, Shai Y
JournalJ Biol Chem
PubMed ID1551864
'Fluorescence resonance energy transfer has been used to study the aggregation and organization of pardaxin and its analogues within lipid membranes. Peptide molecules labeled with 5- (and 6-) carboxyfluorescein at their N-terminal amino acid served as donors in these energy transfer measurements, whereas peptides labeled with 5- (and 6-) carboxytetramethylrhodamine ... More
Kinetic studies by fluorescence resonance energy transfer employing a double-labeled oligonucleotide: hybridization to the oligonucleotide complement and to single-stranded DNA.
AuthorsParkhurst KM, Parkhurst LJ
JournalBiochemistry
PubMed ID7819209
'A single 16-base oligodeoxyribonucleotide was labeled at the 3''-end with fluorescein and at the 5''-end with x-rhodamine (R*oligo*F); the chromophores served as a donor/acceptor pair, respectively, for Förster resonance energy transfer. We exploited the striking differences in the steady-state emission spectra of the R*oligo*F as a single strand and in ... More
Fluorotyping of HLA-A by sequence-specific priming and fluorogenic probing.
AuthorsSlateva K, Camps MA, Blasczyk R
JournalTissue Antigens
PubMed ID9864036
'The aim of our study was to develop a fluorotyping strategy for the HLA-A locus. In contrast to conventional sequence-specific primed PCR (PCR-SSP), which is based on an agarose gel electrophoresis, fluorotyping eliminates the drawback of low sample throughput, low potential for automation and problems related to contamination. Additionally, fluorotyping ... More
AhpF can be dissected into two functional units: tandem repeats of two thioredoxin-like folds in the N-terminus mediate electron transfer from the thioredoxin reductase-like C-terminus to AhpC.
AuthorsPoole LB, Godzik A, Nayeem A, Schmitt JD
JournalBiochemistry
PubMed ID10828978
'AhpF, the flavin-containing component of the Salmonella typhimurium alkyl hydroperoxide reductase system, catalyzes the NADH-dependent reduction of an active-site disulfide bond in the other component, AhpC, which in turn reduces hydroperoxide substrates. The amino acid sequence of the C-terminus of AhpF is 35% identical to that of thioredoxin reductase (TrR) ... More
A three-dimensional model for the hammerhead ribozyme based on fluorescence measurements.
AuthorsTuschl T, Gohlke C, Jovin TM, Westhof E, Eckstein F
JournalScience
PubMed ID7973630
'For the understanding of the catalytic function of the RNA hammerhead ribozyme, a three-dimensional model is essential but neither a crystal nor a solution structure has been available. Fluorescence resonance energy transfer (FRET) was used to study the structure of the ribozyme in solution in order to establish the relative ... More
Enolase exists in the fluid phase of cytoplasm in 3T3 cells.
AuthorsPagliaro L, Kerr K, Taylor DL
JournalJ Cell Sci
PubMed ID2621228
'We have investigated the intracellular distribution and mobility of the glycolytic enzyme enolase, using functional fluorescent analogs labeled with the succinimidyl esters of carboxyfluorescein (F1-enolase) and carboxytetramethylrhodamine (Rh-enolase) In contrast to aldolase, neither native enolase nor labeled enolase gelled filamentous actin (F-actin), as measured by falling-ball viscometry, indicating a lack ... More
Direct measurement of thermodynamic and kinetic parameters of DNA triple helix formation by fluorescence spectroscopy.
AuthorsYang M, Ghosh SS, Millar DP
JournalBiochemistry
PubMed ID7803396
'Direct measurement of thermodynamic and kinetic parameters of oligonucleotide-directed DNA triple helix formation has been achieved by fluorescence spectroscopic methods. Fluorescence resonance energy transfer (FRET) was used to study the binding of an acceptor-labeled single-stranded oligonucleotide to a donor-labeled DNA duplex. Equilibrium binding constants and association rate constants for triplex ... More
A fluorescence anisotropy study of DNA binding by HPV-11 E2C protein: a hierarchy of E2-binding sites.
AuthorsAlexander KA, Phelps WC
JournalBiochemistry
PubMed ID8703960
'Association of the human papillomavirus (HPV) E2 protein with its palindromic DNA-binding site is a necessary step for transcriptional trans-activation. To study the interaction between DNA and E2, the carboxyl-terminal domain of HPV-11 E2 protein (E2C) was expressed in Escherichia coli and purified to homogeneity. The binding affinity of the ... More
Fluorescence characteristics of 5-carboxytetramethylrhodamine linked covalently to the 5' end of oligonucleotides: multiple conformers of single-stranded and double-stranded dye-DNA complexes.
AuthorsVámosi G, Gohlke C, Clegg RM
JournalBiophys J
PubMed ID8842236
'Fluorescence steady-state and lifetime experiments have been carried out on duplex and single-stranded DNA molecules labeled at the 5'' ends with 5-carboxytetramethylrhodamine (TMRh). The temperature and ionic strength of the solutions were varied over large ranges. The results reveal at least three well-defined states of the TMRh-DNA molecules for the ... More
'A substrate for a hypersensitive assay of ribonucleolytic activity was developed in a systematic manner. This substrate is based on the fluorescence quenching of fluorescein held in proximity to rhodamine by a single ribonucleotide embedded within a series of deoxynucleotides. When the substrate is cleaved, the fluorescence of fluorescein is ... More
Chopper, a new death domain of the p75 neurotrophin receptor that mediates rapid neuronal cell death.
AuthorsCoulson EJ, Reid K, Baca M, Shipham KA, Hulett SM, Kilpatrick TJ, Bartlett PF
JournalJ Biol Chem
PubMed ID10882742
'The cytoplasmic juxtamembrane region of the p75 neurotrophin receptor (p75(NTR)) has been found to be necessary and sufficient to initiate neural cell death. The region was named "Chopper" to distinguish it from CD95-like death domains. A 29-amino acid peptide corresponding to the Chopper region induced caspase- and calpain-mediated death in ... More
Simultaneous measurements of actin filament turnover, filament fraction, and monomer diffusion in endothelial cells.
AuthorsMcGrath JL, Tardy Y, Dewey CF, Meister JJ, Hartwig JH
JournalBiophys J
PubMed ID9746549
'The analogous techniques of photoactivation of fluorescence (PAF) and fluorescence recovery after photobleaching (FRAP) have been applied previously to the study of actin dynamics in living cells. Traditionally, separate experiments estimate the mobility of actin monomer or the lifetime of actin filaments. A mathematical description of the dynamics of the ... More
Fluorescence energy transfer dye-labeled primers for DNA sequencing and analysis.
AuthorsJu J, Ruan C, Fuller CW, Glazer AN, Mathies RA
JournalProc Natl Acad Sci U S A
PubMed ID7753809
'Fluorescent dye-labeled DNA primers have been developed that exploit fluorescence energy transfer (ET) to optimize the absorption and emission properties of the label. These primers carry a fluorescein derivative at the 5'' end as a common donor and other fluorescein and rhodamine derivatives attached to a modified thymidine residue within ... More
Regulation of cAMP-dependent protein kinase: enzyme activation without dissociation.
AuthorsYang S, Fletcher WH, Johnson DA
JournalBiochemistry
PubMed ID7756252
'It has become axiomatic that, in contrast to other protein kinases, cAMP-dependent protein kinase (cAPK) is activated only when its catalytic (C) and regulatory (RII2) subunits dissociate. To directly evaluate this postulation, the ability of cAMP to dissociate the holoenzyme form of cAPK was examined by measuring the rotational mobility ... More
GLUT4 and transferrin receptor are differentially sorted along the endocytic pathway in CHO cells.
AuthorsWei ML, Bonzelius F, Scully RM, Kelly RB, Herman GA
JournalJ Cell Biol
PubMed ID9456317
'The trafficking of GLUT4, a facilitative glucose transporter, is examined in transfected CHO cells. In previous work, we expressed GLUT4 in neuroendocrine cells and fibroblasts and found that it was targeted to a population of small vesicles slightly larger than synaptic vesicles (Herman, G.A, F. Bonzelius, A.M. Cieutat, and R.B. ... More
N-type Ca2+ channels are located on somata, dendrites, and a subpopulation of dendritic spines on live hippocampal pyramidal neurons.
AuthorsMills LR, Niesen CE, So AP, Carlen PL, Spigelman I, Jones OT
JournalJ Neurosci
PubMed ID7525892
'In the nervous system the influx of Ca2+ orchestrates multiple biochemical and electrical events essential for development and function. A major route for Ca2+ entry is through voltage-dependent calcium channels (VDCCs). It is becoming increasingly clear that the precise contribution VDCCs make to neuronal function depends not only upon their ... More
Site-specific interaction of thrombin and inhibitors observed by fluorescence correlation spectroscopy.
AuthorsKlingler J, Friedrich T
JournalBiophys J
PubMed ID9336216
'We report on the application of fluorescence correlation spectroscopy (FCS) to observe the interaction between thrombin and thrombin inhibitors. Two site-specific fluorescent labels were used to distinguish between inhibitors directed to the active site, the exosite, or both binding sites of thrombin. For several well-known inhibitors of thrombin, the binding ... More
Fluorescein derivatization of fibrinogen for flow cytometric analysis of fibrinogen binding to platelets.
AuthorsHeilmann E, Hynes LA, Burstein SA, George JN, Dale GL
JournalCytometry
PubMed ID7875035
'Dog and human fibrinogen were derivatized with N-hydroxysuccinimido-fluorescein and utilized for flow cytometric estimation of fibrinogen binding to activated platelets. Fluorescein-fibrinogen binding fulfilled the criteria for specific binding to platelets; the binding was saturable, dependent on agonist activation, and inhibited by unlabeled fibrinogen. In addition, EDTA and barbourin, a KGD-containing ... More
Allosteric effects of a monoclonal antibody against thrombin exosite II.
'We previously isolated a monoclonal antithrombin IgG from a patient with multiple myeloma [Colwell et al. (1997) Br. J. Haematol. 97, 219-226]. Using a panel of 55 surface mutants of recombinant thrombin, we now show that the epitope for the IgG most likely includes Arg-101, Arg-233, and Lys-236 in exosite ... More
Dissociation kinetics of actinomycin D from oligonucleotides with hairpin motifs.
AuthorsChen FM, Jones CM, Johnson QL
JournalBiochemistry
PubMed ID8504076
'The dissociation of 7-aminoactinomycin D (7-AM-ACTD) from d(ATGCATATGCAT), d(ATGCAT-T-ATGCAT), or d(ATGCAT-A-ATGCAT) at 20 degrees C cannot be adequately described by a single-exponential decay and requires a fit with two rate constants. The relative contributions of these two rate processes and their temperature dependence can be attributed to the coexistence of ... More
Membrane interaction and self-assembly within phospholipid membranes of synthetic segments corresponding to the H-5 region of the shaker K+ channel.
AuthorsPeled H, Shai Y
JournalBiochemistry
PubMed ID8347593
'The voltage-activated K+ channels are assumed to be formed by the coassembly of four polypeptide monomers. Each of these monomers is postulated to consist of six transmembrane segments (S1 to S6), and long N- and C-terminal domains. The highly conserved linker, H-5, between the fifth and the sixth transmembrane segments, ... More
Alphabeta protomers of Na+,K+-ATPase from microsomes of duck salt gland are mostly monomeric: formation of higher oligomers does not modify molecular activity.
AuthorsMartin DW, Marecek J, Scarlata S, Sachs JR
JournalProc Natl Acad Sci U S A
PubMed ID10706623
'The distance that separates alphabeta protomers of the Na(+), K(+)-ATPase in microsomes and in purified membranes prepared from duck nasal salt glands was estimated by measuring fluorescence resonance energy transfer between anthroylouabain bound to a population of alphabeta protomers and either N-[7-nitrobenz-2-oxa-1, 3-diazol-4-yl]-6-aminohexyl ouabain or 5-(and-6)-carboxyfluorescein-6-aminohexyl ouabain bound to the ... More
Peptide arrays with designed secondary structures for protein characterization using fluorescent fingerprint patterns.
AuthorsUsui K, Ojima T, Takahashi M, Nokihara K, Mihara H
JournalBiopolymers
PubMed ID15054893
'To realize a practical high-throughput protein-detection system, novel peptide arrays have been constructed using designed peptide libraries with loop, alpha-helix, or beta-strand structures. Here, we describe the overview of the reported designed peptide arrays with loop and alpha-helix structures and the new results of those with beta-strand structures. Initially, several ... More
Molecular determinants of plasma cholesteryl ester transfer protein binding to high density lipoproteins.
'The plasma cholesteryl ester transfer protein (CETP) mediates the transfer of neutral lipids between lipoproteins and is associated with high density lipoproteins (HDL). To understand the mechanism of interaction of CETP with HDL, we studied the binding of pure recombinant CETP to 1-palmitoyl-2-oleoylphosphatidylcholine (POPC)/apoA-I discoidal particles. Separating bound from free ... More
Molecular aptamer beacons for real-time protein recognition.
AuthorsLi JJ, Fang X, Tan W
JournalBiochem Biophys Res Commun
PubMed ID11890667
'One of the most pressing problems facing those attempting to understand the regulation of gene expression and translation is the necessity to monitor protein production in a variety of metabolic states. Thus far, there is no easy solution that will either identify or quantitate proteins in real time. Here we ... More
Intracellular disposition and metabolism of fluorescently-labeled unmodified and modified oligonucleotides microinjected into mammalian cells.
AuthorsFisher TL, Terhorst T, Cao X, Wagner RW
JournalNucleic Acids Res
PubMed ID8396239
'The intracellular distribution and metabolism of microinjected fluorescently-labeled oligonucleotides (ODNs) have been evaluated using confocal fluorescence microscopy. Fluorescent phosphodiester ODNs, microinjected into the cytoplasm of mammalian cells, rapidly accumulate within the nucleus; the fluorescence disappears with a half-life of 15-20 minutes. Microinjected fluorescent phosphorothioate ODNs remain in the nucleus for ... More
Somatodendritic internalization and perinuclear targeting of neurotensin in the mammalian brain.
AuthorsFaure MP, Alonso A, Nouel D, Gaudriault G, Dennis M, Vincent JP, Beaudet A
JournalJ Neurosci
PubMed ID7790901
'Polypeptide hormones and growth factors have long been known to internalize into peripheral target cells as a result of their interaction with cell surface receptors. Studies in culture have suggested that certain neuropeptides might undergo a similar type of translocation in neurons. To investigate this possibility in adult mammalian brain, ... More
High-resolution typing for HLA-DRB1*15 amd -DRB1*16 by fluorescence-marked sequence-specific priming (TaqMan assay).
AuthorsTremmel M, Opelz G, Mytilineos J
JournalTissue Antigens
PubMed ID10599890
'Sequence-specific primed polymerase chain reaction (PCR-SSP) is widely used in HLA laboratories. The TaqMan method, which is described here for high-resolution typing of HLA-DRB1*15 and -DRB1*16, does not require elaborate and time-consuming post-PCR detection steps. In this one-tube assay, conventional PCR-SSP and fluorescence detection of the amplicon with a doubly ... More
Assessment of glutaraldehyde crosslinking efficiency with an amine-specific fluorescent probe.
AuthorsRicks J, Scott M, Vesely I
JournalAnn Thorac Surg
PubMed ID9930456
'BACKGROUND: Crosslinking of heart valves with glutaraldehyde involves the binding of amine groups. We have developed a technique that provides an inverse measure of the degree of tissue fixation by quantifying the amount of unbound amines. METHODS: Whole aortic valves were exposed to 0.5% glutaraldehyde solution for 0, 1, 15, ... More
A new class of homogeneous nucleic acid probes based on specific displacement hybridization.
AuthorsLi Q, Luan G, Guo Q, Liang J
JournalNucleic Acids Res
PubMed ID11788731
'We have developed a new class of probes for homogeneous nucleic acid detection based on the proposed displacement hybridization. Our probes consist of two complementary oligodeoxyribonucleotides of different length labeled with a fluorophore and a quencher in close proximity in the duplex. The probes on their own are quenched, but ... More
Synthesis and pharmacological activity of fluorescent histamine H1 receptor antagonists related to mepyramine.
AuthorsLi L, Kracht J, Peng S, Bernhardt G, Buschauer A
JournalBioorg Med Chem Lett
PubMed ID12657255
'Fluorescently labeled histamine H(1) receptor antagonists were synthesized starting from N-demethylmepyramine by introduction of omega-aminoalkyl chains (2-8 methylene groups in length) followed by derivatization of the terminal NH(2) group with various fluorophores (fluorescein, naphthofluorescein, rhodamine, tetramethylrhodamine, BODIPY, dansyl, and nitrobenzoxadiazole (NBD)). On the isolated guinea pig ileum and in a ... More
Extending the applicability of carboxyfluorescein in solid-phase synthesis.
AuthorsFischer R, Mader O, Jung G, Brock R
JournalBioconjug Chem
PubMed ID12757391
'Optimized coupling protocols are presented for the efficient and automated generation of carboxyfluorescein-labeled peptides. Side products, generated when applying earlier protocols for the in situ activation of carboxyfluorescein, were eliminated by a simple procedure, yielding highly pure fluorescent peptides and minimizing postsynthesis workup. For the cost-efficient labeling of large compound ... More
Importin 13: a novel mediator of nuclear import and export.
AuthorsMingot JM, Kostka S, Kraft R, Hartmann E, Görlich D
JournalEMBO J
PubMed ID11447110
'Importin beta-related receptors mediate translocation through nuclear pore complexes. Co-operation with the RanGTPase system allows them to bind and subsequently release their substrates on opposite sides of the nuclear envelope, which in turn ensures a directed nucleocytoplasmic transport. Here we identify a novel family member from higher eukaryotes that functions ... More
A peptide-based fluorescence resonance energy transfer assay for Bacillus anthracis lethal factor protease.
AuthorsCummings RT, Salowe SP, Cunningham BR, Wiltsie J, Park YW, Sonatore LM, Wisniewski D, Douglas CM, Hermes JD, Scolnick EM
JournalProc Natl Acad Sci U S A
PubMed ID11997440
'A fluorescence resonance energy transfer assay has been developed for monitoring Bacillus anthracis lethal factor (LF) protease activity. A fluorogenic 16-mer peptide based on the known LF protease substrate MEK1 was synthesized and found to be cleaved by the enzyme at the anticipated site. Extension of this work to a ... More
Ultra-high-speed DNA sequencing using capillary electrophoresis chips.
AuthorsWoolley AT, Mathies RA
JournalAnal Chem
PubMed ID8644919
'DNA sequencing has been performed on microfabricated capillary electrophoresis chips. DNA separations were achieved in 50 x 8 microns cross-section channels microfabricated in a 2 in. x 3 in. glass sandwich structure using a denaturing 9% T, 0% C polyacrylamide sieving medium. DNA sequencing fragment ladders were produced and fluorescently ... More
Real-time PCR in the microbiology laboratory.
AuthorsMackay IM
JournalClin Microbiol Infect
PubMed ID15008940
'Use of PCR in the field of molecular diagnostics has increased to the point where it is now accepted as the standard method for detecting nucleic acids from a number of sample and microbial types. However, conventional PCR was already an essential tool in the research laboratory. Real-time PCR has ... More
The physiologic concentration of inositol 1,4,5-trisphosphate in the oocytes of Xenopus laevis.
'To measure the concentration of inositol 1,4,5-trisphosphate ([IP3]) in small regions of single Xenopus oocytes, a biological detector cell was combined with capillary electrophoresis. This method is 10, 000 times more sensitive than all existing assays enabling subcellular measurement of [IP3] in Xenopus oocytes. Upon addition of lysophosphatidic acid to ... More
HEMCAM, an adhesion molecule expressed by c-kit+ hemopoietic progenitors.
AuthorsVainio O, Dunon D, Aïssi F, Dangy JP, McNagny KM, Imhof BA
JournalJ Cell Biol
PubMed ID8978830
'We have characterized the adhesion molecule HEMCAM, which is expressed by hemopoietic progenitors of embryonic bone marrow. HEMCAM belongs to the immunoglobulin superfamily and consists of the V-V-C2-C2-C2 Ig domains. There are three mRNA splice variants. One has a short cytoplasmic tail; another has a long tail; while the third ... More
Optimizing primer--probe design for fluorescent PCR.
AuthorsProudnikov D, Yuferov V, Zhou Y, LaForge KS, Ho A, Kreek MJ
JournalJ Neurosci Methods
PubMed ID12581847
'TaqMan, a variation of fluorescent PCR, is a powerful tool for gene expression and polymorphism studies. Here we describe the design and evaluation of 27 new TaqMan primer-probe sets for rat genes that play a key role in neural signaling. These newly designed and synthesized probes were tested and then ... More
Donor-acceptor distance distributions in a double-labeled fluorescent oligonucleotide both as a single strand and in duplexes.
AuthorsParkhurst KM, Parkhurst LJ
JournalBiochemistry
PubMed ID7819210
'A 16-mer deoxyribonucleotide was labeled at the 5''-end with x-rhodamine and at the 3''-end with fluorescein. Förster resonance energy transfer was used to determine the distribution, P(R), of donor-acceptor distances for the oligomer in three duplex structures (hybridized to complementary strands having 10, 16, and 24 bases) and as a ... More
Antibody-based fluorescence detection of kinase activity on a peptide array.
AuthorsLesaicherre ML, Uttamchandani M, Chen GY, Yao SQ
JournalBioorg Med Chem Lett
PubMed ID12127509
'Peptide-based microarrays allow for high-throughput identification of protein kinase substrates. However, current methods of detecting kinase activity require the use of radioisotopes. We have developed a novel fluorescence-based approach for quantitative detection of peptide phosphorylation on chip using fluorescently-labeled anti-phosphoserine and anti-phosphotyrosine antibodies. This method is sensitive, specific and extremely ... More
Measurement of kinase activation in single mammalian cells.
'We demonstrate a new method for the simultaneous measurement of the activation of key regulatory enzymes within single cells. To illustrate the capabilities of the technique, the activation of protein kinase C (PKC), protein kinase A (PKA), calcium-calmodulin activated kinase II (CamKII), and cdc2 protein kinase (cdc2K) was measured in ... More
A brief survey of methods for preparing protein conjugates with dyes, haptens, and cross-linking reagents.
AuthorsBrinkley M
JournalBioconjug Chem
PubMed ID1616945
Fluorotyping of HLA-C: differential detection of amplicons by sequence-specific priming and fluorogenic probing.
AuthorsLuedeck H, Blasczyk R
JournalTissue Antigens
PubMed ID9458116
'Conventional PCR-SSP, which is based on an agarose gel-based read-out, has the disadvantages of time-consuming post-PCR steps and low potential for automation. The aim of our study was to sort out these drawbacks by establishing a fluorescence-based PCR-SSP system for HLA-C. The assay relies on the sequence-specific identification of amplicons ... More
Self diffusion and spectral modifications of a membrane protein, the Rubrivivax gelatinosus LH2 complex, incorporated into a monoolein cubic phase.
AuthorsTsapis N, Reiss-Husson F, Ober R, Genest M, Hodges RS, Urbach W
JournalBiophys J
PubMed ID11509374
'The light-harvesting complex LH2 from a purple bacterium, Rubrivivax gelatinosus, has been incorporated into the Q230 cubic phase of monoolein. We measured the self-diffusion of LH2 in detergent solution and in the cubic phase by fluorescence recovery after photobleaching. We investigated also the absorption and fluorescence properties of this oligomeric ... More
Simultaneous assay of Src SH3 and SH2 domain binding using different wavelength fluorescence polarization probes.
AuthorsLynch BA, Minor C, Loiacono KA, van Schravendijk MR, Ram MK, Sundaramoorthi R, Adams SE, Phillips T, Holt D, Rickles RJ, MacNeil IA
JournalAnal Biochem
PubMed ID10542110
'pp60(c-src) is a prototypical nonreceptor tyrosine kinase and may play a role in diseases as diverse as cancer and osteoporosis. In Src, the SH3 domain (Src homology 3) binds proteins at specific, proline-rich sequences, while the SH2 domain (Src homology 2) binds phosphotyrosine-containing sequences. Inhibition of Src SH3 and SH2 ... More
Hydrophilic fluorescein derivatives: useful reagents for liposome immunolytic assays.
AuthorsFiechtner M, Wong M, Bieniarz C, Shipchandler MT
JournalAnal Biochem
PubMed ID2817337
'Hydrophilic derivatives of fluorescein containing hydroxyalkyl substituents were synthesized and encapsulated within liposomes. The fluorophores showed significantly more retention with time than did fluorescein, carboxyfluorescein, or calcein. Unlike calcein, the fluorophores are minimally susceptible to fluorescence quenching by Co2+. The utility of these compounds as immunodiagnostic reagents was demonstrated by ... More
Design of a Molecular Beacon DNA Probe with Two Fluorophores This work was partially supported by the Office of Naval Research Young Investigator Award N00014-98-1-0621 and by an NSF Career Award (CHE-9733650). We would like to thank Dr. Richard Hogrefe, Paul Imperial, Kelly Christianson, and the Trilink Oligonucleotide Synthesis group for producing the molecular beacons used in this study.
AuthorsZhang P, Beck T, Tan W
JournalAngew Chem Int Ed Engl
PubMed ID11180338
Fluorescent microtubules break up under illumination.
AuthorsVigers GP, Coue M, McIntosh JR
JournalJ Cell Biol
PubMed ID3417772
'We have synthesized three new fluorescent analogues of tubulin, using fluorescein or rhodamine groups attached to N-hydroxy-succinimidyl esters, and have partially characterized the properties of these analogues. We have also further characterized the tubulin derivatized with dichlorotriazinyl-aminofluorescein that has previously been used in this and other laboratories. Our results show ... More
Transferrin receptor containing the SDYQRL motif of TGN38 causes a reorganization of the recycling compartment but is not targeted to the TGN.
AuthorsJohnson AO, Ghosh RN, Dunn KW, Garippa R, Park J, Mayor S, Maxfield FR, McGraw TE
JournalJ Cell Biol
PubMed ID8991088
'The SDYQRL motif of the cytoplasmic domain of TGN38 is involved in targeting TGN38 from endosomes to the TGN. To create a system for studying this pathway, we replaced the native transferrin receptor (TR) internalization motif (YTRF) with the SDYQRL TGN-targeting motif. The advantages of using TR as a reporter ... More
Factor V Leiden genotyping using real-time fluorescent polymerase chain reaction.
AuthorsSanders Sevall J
JournalMol Cell Probes
PubMed ID10970729
'A fluorogenic probe-based PCR assay (Taqman; Perkin Elmer corp/Applied Biosystems, Foster City, CA, USA) was used for the detection of Factor V Leiden, a point mutation in the factor V gene (G1691A) that is the most common inherited risk factor for Deep Vein Thrombosis. This assay allows for the direct ... More
Structural and kinetic studies of a cisplatin-modified DNA icosamer binding to HMG1 domain B.
AuthorsJamieson ER, Jacobson MP, Barnes CM, Chow CS, Lippard SJ
JournalJ Biol Chem
PubMed ID10212205
'The high mobility group (HMG) domain is a DNA-binding motif found in the non-histone chromosomal proteins, HMG1 and HMG2, and some transcription factors. Experimental evidence has demonstrated that HMG-domain proteins can play a role in sensitizing cells to the anticancer drug cisplatin. Fluorescence resonance energy transfer (FRET) experiments were performed ... More
A novel method for real time quantitative RT-PCR.
AuthorsGibson UE, Heid CA, Williams PM
JournalGenome Res
PubMed ID8908519
'A novel approach to quantitative reverse transcriptase polymerase chain reaction (QC RT-PCR) using real time detection and the 5'' nuclease assay has been developed. Cystic fibrosis transmembrane transductance regulator (CFTR) target mRNA is reverse transcribed, amplified, detected, and quantitated in real time. A fluorogenic probe was designed to detect the ... More
Multiple forms of the 20 S multicatalytic and the 26 S ubiquitin/ATP-dependent proteases from rabbit reticulocyte lysate.
AuthorsHoffman L, Pratt G, Rechsteiner M
JournalJ Biol Chem
PubMed ID1331052
'We have used native gel electrophoresis followed by fluorogenic peptide overlay to identify multiple forms of rabbit reticulocyte multicatalytic protease (MCP) or 20 S protease, and two forms of rabbit 26 S ubiquitin/ATP-dependent protease. An abundant, fast-migrating 20 S complex (20 SF) possesses modest ability to hydrolyze the fluorogenic peptide ... More
IL-6 produced by macrophages infected with Mycobacterium species suppresses T cell responses.
AuthorsVanHeyningen TK, Collins HL, Russell DG
JournalJ Immunol
PubMed ID8977207
'The ability of Mycobacterium bovis Calmette-Guérin bacillus-infected bone marrow-derived macrophages to process and present exogenously added Ags to T cells and stimulate their growth and production of IL-2 was examined. The infected macrophages were inhibited in their ability to activate T cells, and this inhibition could be transferred to uninfected ... More
Visual detection of specific, native interactions between soluble and microbead-tethered alpha-helices from membrane proteins.
AuthorsAshish Wimley WC
JournalBiochemistry
PubMed ID11705363
'Using peptides tethered to polymer microbeads, we have developed a technique for measuring the interactions between the transmembrane alpha-helices of membrane proteins and for screening combinatorial libraries of peptides for members that interact with specific helices from membrane proteins. The method was developed using the well-characterized homodimerization sequence of the ... More
Chemical methods of DNA and RNA fluorescent labeling.
AuthorsProudnikov D, Mirzabekov A
JournalNucleic Acids Res
PubMed ID8948646
'Several procedures have been described for fluorescent labeling of DNA and RNA. They are based on the introduction of aldehyde groups by partial depurination of DNA or oxidation of the 3''-terminal ribonucleoside in RNA by sodium periodate. Fluorescent labels with an attached hydrazine group are efficiently coupled with the aldehyde ... More
Synthesis of fluorescent biochemical tools related to the 2-azetidinone class of cholesterol absorption inhibitors.
AuthorsBurnett DA, Caplen MA, Browne ME, Zhau H, Altmann SW, Davis HR, Clader JW
JournalBioorg Med Chem Lett
PubMed ID11814786
'Fluorescent analogues of the cholesterol absorption inhibitor (CAI), Sch 58235, have been designed and synthesized as single enantiomers. Biological testing reveals that they are potent CAIs and are suitable tools for the investigation of the azetidinone CAI mechanism of action (MOA).' ... More
Microtubule dynamics in living dividing plant cells: confocal imaging of microinjected fluorescent brain tubulin.
AuthorsZhang D, Wadsworth P, Hepler PK
JournalProc Natl Acad Sci U S A
PubMed ID11607116
'Carboxyfluorescein-labeled brain tubulin has been microinjected into stamen hair cells of Tradescantia, and its distribution during mitosis and cytokinesis was examined using confocal laser scanning fluorescence microscopy. The results show that brain tubulin incorporates into plant microtubules and is utilized throughout mitosis and cytokinesis. Microtubule structures that incorporate brain tubulin ... More
Localized measurement of kinase activation in oocytes of Xenopus laevis.
'We have combined a rapid cytoplasmic sampling technique with capillary electrophoresis to measure the activation of protein kinase C (PKC) in a small region (approximately 60 microm) of a Xenopus oocyte. The phosphorylation of a fluorescent PKC substrate was measured following addition of a pharmacological or physiological stimulus to an ... More
Energy-transfer fluorescent reagents for DNA analyses.
AuthorsGlazer AN, Mathies RA
JournalCurr Opin Biotechnol
PubMed ID9013654
'Fluorescence resonance energy transfer has facilitated the development of a new class of high-performance fluorescent labeling reagents for multiplex analyses of nucleic acids. The enhanced emission of energy transfer (ET) primers has provided a decadic improvement in the performance of automated DNA sequencers. The emission spectral purity of ET primers ... More
The Doc toxin and Phd antidote proteins of the bacteriophage P1 plasmid addiction system form a heterotrimeric complex.
AuthorsGazit E, Sauer RT
JournalJ Biol Chem
PubMed ID10358024
'The toxin (Doc) and antidote (Phd) proteins of the plasmid addiction system of bacteriophage P1 were purified as a complex. Cocrystals of the complex contained a 2:1 molar ratio of Phd:Doc as assayed by dye binding following SDS-polyacrylamide gel electrophoresis and as determined by amino acid analysis. Gel filtration and ... More
Structural basis for PRYSPRY-mediated tripartite motif (TRIM) protein function.
AuthorsJames LC, Keeble AH, Khan Z, Rhodes DA, Trowsdale J
JournalProc Natl Acad Sci U S A
PubMed ID17400754
'The human tripartite motif (TRIM) family comprises 70 members, including HIV restriction factor TRIM5alpha and disease-associated proteins TRIM20 (pyrin) and TRIM21. TRIM proteins have conserved domain architecture but diverse cellular roles. Here, we describe how the C-terminal PRYSPRY domain mediates diverse TRIM functions. The crystal structure of TRIM21 PRYSPRY in ... More
Energy transfer primers with 5- or 6-carboxyrhodamine-6G as acceptor chromophores.
AuthorsHung SC, Ju J, Mathies RA, Glazer AN
JournalAnal Biochem
PubMed ID8660606
'Energy-transfer (ET) fluorescent primers for DNA sequencing and multiplex genetic analysis (Ju, J., Ruan, C., Fuller, C. W., Glazer, A. N., and Mathies, R. A. (1995) Proc. Natl. Acad. Sci. USA 92, 4347-4351) are named according to the convention D-N-A, where D is the donor, N is the number of ... More
Quantitation of fluorescent nucleotide incorporation by capillary gel electrophoresis and laser-induced fluorescence detection.
AuthorsCruickshank KA
JournalAnal Biochem
PubMed ID10094771
'A method has been developed by which enzymatically incorporated fluorophore-labeled nucleotide sites in nucleic acid can be quantitated by degradation of nanogram quantities of DNA followed by capillary gel electrophoretic analysis with fluorescence detection. In this way the differing relative labeling densities achieved using either C5-substituted dUTP''s or N4-substituted dCTP''s ... More
Formation and dissociation of short-lived class II MHC-peptide complexes.
AuthorsWitt SN, McConnell HM
JournalBiochemistry
PubMed ID8110789
'The incubation of a detergent-solubilized class II MHC protein with excess peptide at 37 degrees C leads to the formation of long-lived protein-peptide complexes (alpha beta P*), which have reported dissociation half-times at 37 degrees C from 30 to 100 h (alpha beta P*-->alpha beta + P*). Here we report ... More
Fluorescent labeling of cell-free synthesized proteins by incorporation of fluorophore-conjugated nonnatural amino acids.
AuthorsKang SH, Jun SY, Kim DM
JournalAnal Biochem
PubMed ID17113028
'Although fluorescent dyes, such as fluorescein derivatives, have bulky and complex structures, nonnatural amino acids carrying these fluorescein derivatives are acceptable by the Escherichia coli ribosome and are useful for the cotranslational fluorescent labeling of cell-free synthesized proteins. Surprisingly, the incorporation efficiency of nonnatural amino acids carrying fluorescein derivatives into ... More
Transferrin-binding protein complex is the receptor for transferrin uptake in Trypanosoma brucei.
AuthorsSteverding D, Stierhof YD, Fuchs H, Tauber R, Overath P
JournalJ Cell Biol
PubMed ID8522581
'In Trypanosoma brucei, the products of two genes, ESAG 6 and ESAG 7, located upstream of the variant surface glycoprotein gene in a polycistronic expression site form a glycosylphosphatidylinositol-anchored transferrin-binding protein (TFBP) complex. It is shown by gel filtration and membrane-binding experiments that the TFBP complex is heterodimeric and binds ... More
Single-cell analysis of T-cell responses in vitro and in vivo: role of costimulatory signals.
AuthorsWells A, Gudmundsdottir H, Walsh M, Turka LA
JournalTransplant Proc
PubMed ID10083353
Triple fluorescence energy transfer in covalently trichromophore-labeled DNA.
AuthorsTong AK, Jockusch S, Li Z, Zhu HR, Akins DL, Turro NJ, Ju J
JournalJ Am Chem Soc
PubMed ID11749560
Simultaneous detection of TaqMan probes containing Fam and Tamra reporter fluorophores.
AuthorsNasarabadi S, Milanovich F, Richards J, Belgrader P
JournalBiotechniques
PubMed ID10631486
4',5'-Dimethoxy-6-carboxyfluorescein: a novel dipole-dipole coupled fluorescence energy transfer acceptor useful for fluorescence immunoassays.
AuthorsKhanna PL, Ullman EF
JournalAnal Biochem
PubMed ID7006452
A DNA assay based on fluorescence resonance energy transfer and DNA triplex formation.
AuthorsYang M, Ren LQ, Huang M, Kong RY, Fong WF
JournalAnal Biochem
PubMed ID9618207
Preparation of modified tubulins.
AuthorsHyman A, Drechsel D, Kellogg D, Salser S, Sawin K, Steffen P, Wordeman L, Mitchison T
JournalMethods Enzymol
PubMed ID2034137
Protein labeling with fluorescent probes.
AuthorsHolmes KL, Lantz LM
JournalMethods Cell Biol
PubMed ID11060842
A new, highly sensitive method for the detection and quantification of penicillin-binding proteins.
AuthorsGalleni M, Lakaye B, Lepage S, Jamin M, Thamm I, Joris B, Frère JM
JournalBiochem J
PubMed ID8385929
A new method for the identification and quantification of penicillin-binding proteins is described which uses fluorescein-coupled penicillins. It allows the rapid detection of 0.2 pmol with the naked eye and 2 fmol with the help of an A.L.F. automatic DNA sequencer. Direct labelling can also be performed on whole bacterial ... More
Antibody-antigen binding constants determined in solution-phase with the threshold membrane-capture system: binding constants for anti-fluorescein, anti-saxitoxin, and anti-ricin antibodies.
AuthorsDill K, Lin M, Poteras C, Fraser C, Hafeman DG, Owicki JC, Olson JD
JournalAnal Biochem
PubMed ID8203727
Affinities of various monoclonal and polyclonal antibodies for fluorescein-containing antigens, saxitoxin and ricin, were determined by using a light addressable potentiometric sensor-based system (Threshold). The dissociation constants, determined from Scatchard plots, ranged from 2 x 10(-7) to approximately 3 x 10(-12) M. Dissociation constants for fluorescein and saxitoxin were compared ... More
Modeling the kinetics of acylation of insulin using a recursive method for solving the systems of coupled differential equations.
AuthorsGrzybowski BA, Anderson JR, Colton I, Brittain ST, Shakhnovich EI, Whitesides GM
JournalBiophys J
PubMed ID10653778
This paper describes a theoretical method for solving systems of coupled differential equations that describe the kinetics of complicated reaction networks in which a molecule having multiple reaction sites reacts irreversibly with multiple equivalents of a ligand (reagent). The members of the network differ in the number of equivalents of ... More
Multiplex capillary denaturing high-performance liquid chromatography with laser-induced fluorescence detection.
AuthorsXiao W, Stern D, Jain M, Huber CG, Oefner PJ
JournalBiotechniques
PubMed ID11414227
Denaturing high-performance liquid chromatography (DHPLC) is a sensitive, robust, and operationally inexpensive method for the detection of single-base substitutions and small deletions and insertions. To increase sample throughout, we have developed a multiplexing strategy using fluorophores to distinguish different PCR products. The system combines recent advances in the synthesis of ... More
Synthesis, purification and kinetic properties of fluorescein-labelled penicillins.
AuthorsLakaye B, Damblon C, Jamin M, Galleni M, Lepage S, Joris B, Marchand-Brynaert J, Frydrych C, Frere JM
JournalBiochem J
PubMed ID8198525
The synthesis and properties of six fluorescein-labelled penicillins are reported. The two isomers of fluoresceyl-glycyl-6-amino-penicillanic acid are probably the best compounds to use for detection of all the penicillin-binding proteins (PBPs) present in a bacterial membrane preparation. However, the derivatives of ampicillin were much more efficient against Enterobacter aerogenes PBP3. ... More
Light-emitting properties of recombinant semi-synthetic aequorins and recombinant fluorescein-conjugated aequorin for measuring cellular calcium.
AuthorsShimomura O, Musicki B, Kishi Y, Inouye S
JournalCell Calcium
PubMed ID8519061
15 kinds of recombinant semi-synthetic aequorins and a recombinant fluorescein-conjugated aequorin were prepared and their properties in Ca(2+)-triggered luminescence were studied. The semi-synthetic aequorins showed a wide range of Ca(2+)-sensitivity. The luminescence intensity of a high-sensitivity type (hcp-aequorin) was greater than 10(4)-times that of a low-sensitivity type (n-aequorin) at pCa ... More
Stress renders T cell blasts sensitive to killing by activated syngeneic NK cells.
AuthorsRabinovich BA, Shannon J, Su RC, Miller RG
JournalJ Immunol
PubMed ID10946262
Exposure of primary T cell blasts to stress in the forms of heat, hydrogen peroxide, or high-density growth conditions resulted in a state of enhanced susceptibility to killing by syngeneic IL-2-activated NK cells or lymphokine-activated killer cells, but not to killing by CTL. Cytotoxicity was perforin mediated and was not ... More
Characterization of protein-hapten conjugates. 2. Electrospray mass spectrometry of bovine serum albumin-hapten conjugates.
AuthorsAdamczyk M, Gebler JC, Mattingly PG
JournalBioconjug Chem
PubMed ID8853461
Fifteen hapten-bovine serum albumin (BSA) conjugates were prepared from five commercially available activated haptens. Each hapten was coupled to BSA at three different ratios. The conjugates were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two mass spectrometry (MS) methods: matrix-assisted laser desorption ionization (MALDI) and liquid chromatography-electrospray ... More
Biotin-fluorophore conjugates with poly(ethylene glycol) spacers retain intense fluorescence after binding to avidin and streptavidin.
AuthorsGruber HJ, Marek M, Schindler H, Kaiser K
JournalBioconjug Chem
PubMed ID9258455
Conventional biotin-fluorophore conjugates with approximately 14 atom spacers lose most of their fluorescence when binding to avidin or streptavidin, as is demonstrated in the present study. This explains the unusual fact that only biotinylated marker enzymes, but not fluorescent biotins, are regularly used in bioanalytic assays. Novel biotin-spacer-fluorophore conjugates are ... More
Reduction of the rate of fluorescence decay of FITC- and carboxyfluorescein-stained cells by anti-FITC antibodies.
AuthorsAbuknesha RA, al-Mazeedi HM, Price RG
JournalHistochem J
PubMed ID1577624
Anti-fluorescein antibodies were found to prevent the fading of emitted fluorescence from fibroblasts stained with fluorescein-labelled fibronectin antibodies. The prevention of fading is the result of specific binding of the fluorochromes present on the stained cells by the anti-fluorescein antibodies. The sheep anti-FITC antibody used in this study was equally ... More
Nonradioactive labeling of RNA.
AuthorsIgloi GL
JournalAnal Biochem
PubMed ID8789156
A modified ribodinucleotide (named pSEEp) has been synthesized using commercially available components on a DNA synthesizer. The presence of a 3'-terminal primary amino group permits its coupling to a range of nonradioactive labels, exemplified here by fluorescein. The product of chemical derivatization of the parent dinucleotide is a good substrate ... More
Prevention of atherosclerosis in apolipoprotein E-deficient mice by bone marrow transplantation.
AuthorsLinton MF, Atkinson JB, Fazio S
JournalScience
PubMed ID7863332
Apolipoprotein E (apoE) deficiency causes severe hyperlipidemia and atherosclerosis in humans and in gene-targeted mice. Although the majority of apoE in plasma is of hepatic origin, apoE is synthesized by a variety of cell types, including macrophages. Because macrophages derive from hematopoietic cells, bone marrow transplantation was used to examine ... More
Typing of the short tandem repeat D8S347 locus with different fluorescence markers.
AuthorsPöltl R, Luckenbach C, Fimmers R, Ritter H
JournalElectrophoresis
PubMed ID9504824
The short tandem repeat (STR) locus D8S347 was analyzed by capillary electrophoresis. Sequencing data and a population study of 203 individuals from a southwestern German population are presented. We detected 12 different alleles, 340-388 bp in length, and found 40 different genotypes. The heterozygosity index was 85.7%. Futhermore, we investigated ... More
Antibody microarray-based profiling of complex specimens: systematic evaluation of labeling strategies.
Antibody microarrays have often had limited success in detection of low abundant proteins in complex specimens. Signal amplification systems improve this situation, but still are quite laborious and expensive. However, the issue of sensitivity is more likely a matter of kinetically appropriate microarray design as demonstrated previously. Hence, we re-examined ... More
FRET and competing processes between conjugated polymer and dye substituted DNA strands: a comparative study of probe selection in DNA detection.
AuthorsAl Attar HA, Monkman AP,
JournalBiomacromolecules
PubMed ID19334782
Fluorescence resonance energy transfer (FRET) between water-soluble conjugated polymer, poly-(9,9-bis(6'-N,N,N-trimethylammonium)-hexyl-fluorene phenylene) bromide, and ssDNA's labeled with four different types of dyes (Pacific-blue, Alexa-fluor 430, Fluorescein, and ROX) has been investigated. The effect of spectral overlap and Stokes-shift on the efficiency and properties of FRET were studied. In the DNA sequence ... More