Biotin binding to avidin. Oligosaccharide side chain not required for ligand association.
AuthorsHiller Y, Gershoni JM, Bayer EA, Wilchek M
JournalBiochem J
PubMed ID3435435
A commercially available, purified preparation of avidin was found to comprise two polypeptide bands (Mr 18,000 and Mr 15,500 respectively). Both bands bound biotin as assessed by biotin overlays of protein blots. The Mr 15,500 polypeptide was found to differ from the Mr 18,000 polypeptide only in its sugar content. ... More
Electron microscopy of complexes of isolated acetylcholine receptor, biotinyl-toxin, and avidin.
AuthorsHoltzman E, Wise D, Wall J, Karlin A
JournalProc Natl Acad Sci U S A
PubMed ID6952187
The principal curarimimetic toxin of Naja naja siamensis derivatized with biotinyl groups binds specifically both to acetylcholine receptor, isolated from Torpedo californica electric tissue, and to avidin. Isolated complexes of receptor monomer or dimer, biotinyl-toxin, and avidin were negatively stained and examined in the scanning transmission electron microscope. We measured ... 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
Development of homogeneous binding assays based on fluorescence resonance energy transfer between quantum dots and Alexa Fluor fluorophores.
AuthorsNikiforov TT, Beechem JM
JournalAnal Biochem
PubMed ID16860286
'We studied the fluorescence resonance energy transfer (FRET) between quantum dots emitting at 565, 605, and 655 nm as energy donors and Alexa Fluor fluorophores with absorbance maxima at 594, 633, 647, and 680 nm as energy acceptors. As a first step, we prepared covalent conjugates between all three types ... More
The FITC-anti-FITC system is a sensitive alternative to biotin-streptavidin in ELISA.
AuthorsHarmer IJ, Samuel D
JournalJ Immunol Methods
PubMed ID2503563
'In a model ELISA system alkaline phosphatase (AP) absorbed onto microtitre wells was employed as the target antigen. The antigen was then reacted with a monoclonal antibody to AP either unlabelled or labelled with (a) FITC and (b) biotin. The bound anti-AP was then detected with horseradish peroxidase (HRP) conjugates ... More
The association of CRKII with C3G can be regulated by integrins and defines a novel means to regulate the mitogen-activated protein kinases.
AuthorsBuensuceso CS, O'Toole TE
JournalJ Biol Chem
PubMed ID10777617
'In studies to define mechanisms of ERK activation in Chinese hamster ovary cells, we have observed an inverse correlation between CRKII-C3G complex formation and ERK activity. That is, we were able to coprecipitate the guanine nucleotide exchange factor C3G with the adaptor protein CRKII in lysates from suspended cells that ... More
Basic studies on heterobifunctional biotin-PEG conjugates with a 3-(4-pyridyldithio)propionyl marker on the second terminus.
AuthorsKaiser K, Marek M, Haselgrübler T, Schindler H, Gruber HJ
JournalBioconjug Chem
PubMed ID9258454
'Heterobifunctional poly(ethylene glycol) (PEG) derivatives with a biotin terminus have been synthesized and characterized with respect to avidin binding. Unambiguous measurement of biotinyl and pyridyldithiopropionyl end groups was established by selecting suitable assays and introducing necessary modifications. Functional studies on the binding of biotin-PEG conjugates to avidin tetramers revealed much ... More
Epidermal growth factor-stimulated calcium ion transients in individual A431 cells: initiation kinetics and ligand concentration dependence.
AuthorsCheyette TE, Gross DJ
JournalCell Regul
PubMed ID1801919
'The A431 epidermoid carcinoma cell line responds to epidermal growth factor (EGF) stimulation with a number of rapid changes, including alterations in free cytosolic calcium ion concentration ([Ca2+]i). At the single cell level, these changes in [Ca2+]i are known to proceed after a clear lag phase subsequent to EGF stimulus ... More
Blockade of the voltage-gated potassium channel Kv1.3 inhibits immune responses in vivo.
AuthorsKoo GC, Blake JT, Talento A, Nguyen M, Lin S, Sirotina A, Shah K, Mulvany K, Hora D, Cunningham P, Wunderler DL, McManus OB, Slaughter R, Bugianesi R, Felix J, Garcia M, Williamson J, Kaczorowski G, Sigal NH, Springer MS, Feeney W
JournalJ Immunol
PubMed ID9164927
'The voltage activated K+ channel (Kv1.3) has recently been identified as the molecule that sets the resting membrane potential of peripheral human T lymphoid cells. In vitro studies indicate that blockage of Kv1.3 inhibits T cell activation, suggesting that Kv1.3 may be a target for immunosuppression. However, despite the in ... More
On the mode of liposome-cell interactions. Biotin-conjugated lipids as ultrastructural probes.
AuthorsBayer EA, Rivnay B, Skutelsky E
JournalBiochim Biophys Acta
PubMed ID420828
'An efficient method for labeling and visualizing phospholipids at the ultrastructural level is described. Biotin was coupled to the amines of appropriate phospholipids via the N-hydroxysuccinimide ester. The biotinylated lipid could be specifically labeled by ferritin-avidin conjugates and detected by transmission electron microscopy. The lipid derivatives were analyzed and evaluated ... More
The immunomodulatory beta-galactoside-specific lectin from mistletoe: partial sequence analysis, cell and tissue binding, and impact on intracellular biosignalling of monocytic leukemia cells.
AuthorsGabius HJ, Walzel H, Joshi SS, Kruip J, Kojima S, Gerke V, Kratzin H, Gabius S
JournalAnticancer Res
PubMed ID1622124
'Nanogram quantities of the beta-galactoside-specific lectin from mistletoe (ML-I) that is composed of two different types of subunits exhibit immunomodulatory potency and enhance cytokine secretion in vitro and in vivo. Partial sequence analysis of the carbohydrate-binding B chain revealed a ragged N-terminus and overall homologies to the B subunit of ... More
Avidin attachment to red blood cells via a phospholipid derivative of biotin provides complement-resistant immunoerythrocytes.
AuthorsMuzykantov VR, Smirnov MD, Klibanov AL
JournalJ Immunol Methods
PubMed ID8429223
'Preincubation of red blood cells (RBC) in an aqueous dispersion of biotin-phosphatidylethanolamine (biotin-PE) provides binding sites for avidin on the surface of these cells (up to 5 x 10(5) avidin molecules per cell). Previously we have shown that biotin covalently attached to the surface of RBC by a chemical reaction ... More
Slow and fast myosin heavy chain content defines three types of myotubes in early muscle cell cultures.
AuthorsMiller JB, Crow MT, Stockdale FE
JournalJ Cell Biol
PubMed ID3902852
'We prepared monoclonal antibodies specific for fast or slow classes of myosin heavy chain isoforms in the chicken and used them to probe myosin expression in cultures of myotubes derived from embryonic chicken myoblasts. Myosin heavy chain expression was assayed by gel electrophoresis and immunoblotting of extracted myosin and by ... More
Continuous-flow, on-line monitoring of biospecific interactions using electrospray mass spectrometry.
AuthorsHogenboom AC, de Boer AR, Derks RJ, Irth H
JournalAnal Chem
PubMed ID11534702
'A continuous-flow analytical screening system is presented using electrospray mass spectrometry to measure the interaction of biologically active compounds with soluble affinity proteins. The biochemical detection system is based on a solution-phase, homogeneous assay. In a first step, compounds to be screened (e.g., biotinylated compounds, concentration range 10-1,000 nmol/L) are ... More
Movement of myosin-coated beads on oriented filaments reconstituted from purified actin.
AuthorsSpudich JA, Kron SJ, Sheetz MP
JournalNature
PubMed ID3925346
'Although the biochemical properties of the actin/myosin interaction have been studied extensively using actin activation of myosin ATPase as an assay, until recently no well-defined assay has been available to measure the mechanical properties of ATP-dependent movement of myosin along actin filaments. The first direct measurements of the rate of ... More
A rapid and sensitive non-competitive avidin-biotin assay for lactoferrin.
AuthorsVilja P, Krohn K, Tuohimaa P
JournalJ Immunol Methods
PubMed ID3881534
'We have developed an avidin-biotin assay for the detection of lactoferrin in human seminal plasma. We compared 5 modifications of this assay, and found the non-competitive avidin-biotin assay (NABA) to be the most sensitive. The detection limit of 3-step NABA was 0.2 or 0.1 ng lactoferrin/ml, depending whether avidin-biotin-peroxidase complex ... More
Expression of Lyt-1 by a subset of B lymphocytes.
AuthorsManohar V, Brown E, Leiserson WM, Chused TM
JournalJ Immunol
PubMed ID6177768
'Using two-color flow cytometry and multiparameter data analysis, we have shown that the IgM bright, large subset of mouse splenic B lymphocytes express Lyt-1. This is not due to B cell uptake of immune complexes of Lyt-1 and antibody from T cells. The IgM bright cells of autoimmune NZB mice ... More
Biotinylated erythrocytes: in vivo survival and in vitro recovery.
AuthorsSuzuki T, Dale GL
JournalBlood
PubMed ID3304462
'Rabbit erythrocytes were biotinylated by reaction with N-hydroxysuccinimidobiotin; the average level of biotinylation was 25,000 molecules per erythrocyte. These biotinylated cells exhibited a normal survival rate when reinfused into rabbits. Two studies demonstrated that the biotin label was stable in vivo. The first was a double-labeling experiment where the biotinylated ... More
Erratic deposition of agrin during the formation of Xenopus neuromuscular junctions in culture.
AuthorsAnderson MJ, Shi ZQ, Grawel R, Zackson SL
JournalDev Biol
PubMed ID7601300
'In order to disclose the mechanism that regulate synapse development we compared the distributions of agrin, acetylcholine receptors (AChR) and a basal lamina heparan sulfate proteoglycan (HSPG) in sections and cultures prepared from Xenopus laevis and Ambystoma mexicanum embryos. While agrin, AChR and HSPG may accumulate almost synchronously at synapses ... More
Quantitative analysis of Fc gamma receptors on murine spleen cell populations by using dual parameter flow cytometry.
AuthorsTitus JA, Finkelman FD, Stephany DA, Jones JF, Segal DM
JournalJ Immunol
PubMed ID6234350
'The expression of Fc gamma R on subsets of mouse spleen cells was examined by dual parameter flow microfluorometry. B cells were detected by labeling them with antibodies against sIgM, sIgD, sIgG, or I-A; essentially all B cells expressed Fc gamma R. The number of Fc gamma R per cell ... More
Use of the avidin-biotin complex for specific staining of biological membranes in electron microscopy.
AuthorsHeitzmann H, Richards FM
JournalProc Natl Acad Sci U S A
PubMed ID4139715
'To expand the electron microscopist''s options in localization and visualization, a new and general staining technique has been tested. The avidin-biotin complex serves as a coupling between the electron-dense marker, ferritin, and points of interest in biological samples. When specific cellular components are tagged with biotin, those components may be ... More
A comparison of non-radioisotopic hybridization assay methods using fluorescent, chemiluminescent and enzyme labeled synthetic oligodeoxyribonucleotide probes.
AuthorsUrdea MS, Warner BD, Running JA, Stempien M, Clyne J, Horn T
JournalNucleic Acids Res
PubMed ID3387214
'N4-[N-(6-trifluoroacetylamidocaproyl)-2-aminoethyl]-5''-O-dimethoxy trityl -5-methyl-2''-deoxycytidine-3''-N,N-diisopropyl-methylphosphoramidite++ + has been synthesized. This N4-alkylamino deoxycytidine derivative has been incorporated into oligonucleotide probes during chemical DNA synthesis. Subsequent to deprotection and purification, fluorescent (fluorescein, Texas Red and rhodamine), chemiluminescent (isoluminol), and enzyme (horseradish peroxidase, alkaline phosphatase) labels have been specifically incorporated. Detection limits of the labels ... More
Anterograde tracing of retinal axons in the avian embryo with low molecular weight derivatives of biotin.
AuthorsHalfter W
JournalDev Biol
PubMed ID2433173
'Several reactive biotin esters were injected into the eyes of chick and quail embryos at various stages of development. Four of the biotin esters reacted with molecules of the eye tissue and were detected with light and electron microscopy in fluorescein isothiocyanate and peroxidase-avidin incubated sections and whole mounts. Intra ... More
Selective labeling of functional groups on membrane proteins or glycoproteins using reactive biotin derivatives and 125I-streptavidin.
AuthorsRoffman E, Meromsky L, Ben-Hur H, Bayer EA, Wilchek M
JournalBiochem Biophys Res Commun
PubMed ID3085669
'Amino groups, sulfhydryl groups or oxidation-induced aldehydes on erythrocyte membrane proteins and/or glycoproteins, were reacted with biotinyl N-hydroxysuccinimide ester (BNHS), 3-(N-maleimido-propionyl) biocytin (MPB) or biocytin hydrazide (BCHZ), respectively. The detergent-lysed biotinylated samples were subjected to SDS-polyacrylamide gel electrophoresis, and the proteins were transferred onto nitrocellulose membranes. The blot was then ... More
Quantification of effector/target conjugation involving natural killer (NK) or lymphokine activated killer (LAK) cells by two-color flow cytometry.
'Precise estimates of the frequency of NK- and LAK-target conjugates were obtained by two-color flow cytometry using hydroethidine and calcein as intracellular labels for target cells and effector cells, respectively. These two dyes can easily be used with a standard single-laser flow cytometer with excellent signal separation and dye retention. ... More
Concurrent measurement of the survival of two populations of rabbit platelets labeled with either two PKH lipophilic dyes or two concentrations of biotin.
AuthorsRand ML, Wang H, Mody M, Chu I, Treutiger I, Nguyen A, Packham MA, Freedman J
JournalCytometry
PubMed ID11813201
'BACKGROUND: To avoid radioisotopic labeling and permit comparison of the survival of two platelet populations concurrently in one animal, we compared simultaneous recoveries and survival times of homologous rabbit platelets labeled in vitro with the lipophilic dyes PKH26 (red fluorescing) and PKH67 (green fluorescing) and with two levels of biotin ... More
High-purity isolation of bullfrog hair bundles and subcellular and topological localization of constituent proteins.
AuthorsGillespie PG, Hudspeth AJ
JournalJ Cell Biol
PubMed ID1704375
'The small number of hair cells in auditory and vestibular organs severely impedes the biochemical characterization of the proteins involved in mechano-electrical transduction. By developing an efficient and clean "twist-off" method of hair bundle isolation, and by devising a sensitive, nonradioactive method to detect minute quantities of protein, we have ... 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
A brief survey of methods for preparing protein conjugates with dyes, haptens, and cross-linking reagents.
AuthorsBrinkley M
JournalBioconjug Chem
PubMed ID1616945
Purification and characterization of membrane proteins.
AuthorsKraehenbuhl JP, Bonnard C
JournalMethods Enzymol
PubMed ID2167430
Preparation and properties of anti-biotin antibodies.
AuthorsKohen F, Bagci H, Barnard G, Bayer EA, Gayer B, Schindler DG, Ainbinder E, Wilchek M
JournalMethods Enzymol
PubMed ID9211297
Use of the avidin-biotin complex for the localization of actin and myosin with fluorescence microscopy.
AuthorsHeggeness MH, Ash JF
JournalJ Cell Biol
PubMed ID326797
'A new indirect method for fluorescence localization of proteins making use of the avidin-biotin complex is described. We have prepared both a biotin-modified rabbit heavy meromyosin (BHMM) and a biotin-modified antibody to a smooth muscle myosin. After fixation, cells can be treated with either BHMM, which binds to actin, or ... More
Non-radioactive labelling and immunoprecipitation analysis of leukocyte surface proteins using different methods of protein biotinylation.
AuthorsKähne T, Ansorge S
JournalJ Immunol Methods
PubMed ID8308295
'The biotinylation of surface proteins and the detection of immunoprecipitated protein(s) after transfer to nitrocellulose using chemiluminescence methods is a highly sensitive alternative to hazardous radioactive labelling procedures. Ligation of proteins using the common biotin-NHS-ester (N-hydroxysuccinimido-biotin) is often associated with a decrease in immunoreactivity. Here a new non-radioactive method of ... More
The preparation and application of functionalised synthetic oligonucleotides: III. Use of H-phosphonate derivatives of protected amino-hexanol and mercapto-propanol or -hexanol.
AuthorsSinha ND, Cook RM
JournalNucleic Acids Res
PubMed ID3362678
'Syntheses of H-phosphonate salts (4a-e) of N/S-protected alcohols such as 6-aminohexan-1-ol, 3-mercaptopropan-1-ol and 6-mercaptohexan-1-ol are described using 2-chloro-5,6-benzo-1,3,2-phosphorin-4-one (2) as the phosphonylating agent. The H-phosphonate salts (4a-e), in the presence of pivaloyl chloride or adamantoyl chloride as an activator, were coupled to the 5''-end of synthetic oligonucleotides on solid supports ... More
Protein blotting: detection of proteins with colloidal gold, and of glycoproteins and lectins with biotin-conjugated and enzyme probes.
AuthorsRohringer R, Holden DW
JournalAnal Biochem
PubMed ID2984955
'Methods to detect "native" proteins immobilized on nitrocellulose membranes in spot tests or on blots prepared from polyacrylamide slab gels after electrophoretic separation are described. Gold sols were found to be useful as general stains for proteins: They are polychromatic, yield an indelible record, and are complementary to india ink ... More
Cellular localization of GM1-ganglioside with biotinylated choleragen and avidin peroxidase in primary cultured cells from rat brain.
AuthorsAsou H, Brunngraber EG, Jeng I
JournalJ Histochem Cytochem
PubMed ID6195214
'A new technique capable of demonstrating the presence and cellular localization of the ganglioside GM1 in primary cultured cells from the brains of newborn rats is described. The method is based on the highly specific binding of biotinylated choleragen to ganglioside GM1, and takes advantage of the high affinity of ... More
Synthesis and use of labelled nucleoside phosphoramidite building blocks bearing a reporter group: biotinyl, dinitrophenyl, pyrenyl and dansyl.
AuthorsRoget A, Bazin H, Teoule R
JournalNucleic Acids Res
PubMed ID2798121
'The synthesis of protected nucleoside phosphoramidites bearing various markers such as biotinyl, dinitrophenyl, dansyl and pyrenyl groups are reported. These labelled deoxynucleosides phosphoramidites were used for solid phase oligonucleotide synthesis in the same way than the usual protected phosphoramidities without any change in the synthetic cycle and the deprotection step. ... More
Avidin-HRP conjugates in biotin-avidin immunoenzyme cytochemistry.
AuthorsBoorsma DM, Van Bommel J, Vanden Heuvel J
JournalHistochemistry
PubMed ID3013808
'Avidin-HRP conjugates were prepared, analysed and tested for avidin-biotin immunocytochemistry. Suitable biotinylation of enzymes, antigens and antibody was obtained by reacting biotin at equimolar ratio to epsilon aminogroups in proteins. The avidin-biotin interaction was used for immunocytochemical detection of phenomena in the field of immunology, i.e. immune complex trapping, specific ... More
Biotin and fluorescent labeling of RNA using T4 RNA ligase.
AuthorsRichardson RW, Gumport RI
JournalNucleic Acids Res
PubMed ID6194506
'Biotin, fluorescein, and tetramethylrhodamine derivatives of P1-(6-aminohex-1-yl)-P2-(5''-adenosine) pyrophosphate were synthesized and used as substrates with T4 RNA ligase. In the absence of ATP, the non-adenylyl portion of these substrates is transferred to the 3''-hydroxyl of an RNA acceptor to form a phosphodiester bond and the AMP portion is released. E. ... More
The use of amphipols as universal molecular adapters to immobilize membrane proteins onto solid supports.
'Because of the importance of their physiological functions, cell membranes represent critical targets in biological research. Membrane proteins, which make up approximately 1/3 of the proteome, interact with a wide range of small ligands and macromolecular partners as well as with foreign molecules such as synthetic drugs, antibodies, toxins, or ... More
Novel biotinylated nucleotide--analogs for labeling and colorimetric detection of DNA.
AuthorsGebeyehu G, Rao PY, SooChan P, Simms DA, Klevan L
JournalNucleic Acids Res
PubMed ID3588302
'A series of dATP and dCTP nucleotide analogs have been synthesized which are modified by attachment of aliphatic linkers containing a functional group to the amino-nitrogen at the hydrogen bonding positions of the bases, that is, at the 6-position of adenine and the 4-position of cytosine. These nucleotides are incorporated ... More
Effective avidin coating of erythrocytes by disulfide bond formation. Production of a stable rosetting agent for multiple applications.
AuthorsFürfang J, Thierfelder S
JournalJ Immunol Methods
PubMed ID3522744
'A method is described for covalently coupling avidin to the membrane of glutaraldehyde-treated erythrocytes. The method is based on the use of the heterobifunctional coupling reagent N-succinimidyl-3-(2-pyridyldithio)propionate (SPDP) and appears to be more effective than other protein coupling techniques. The resulting avidin-coated red cells are very stable and can be ... More
Up-regulation of interleukin 4/B-cell stimulatory factor 1 receptor expression.
AuthorsOhara J, Paul WE
JournalProc Natl Acad Sci U S A
PubMed ID3263648
'The expression of interleukin 4 (IL-4) receptors on resting T and B lymphocytes was enhanced 4- to 8-fold by IL-4 stimulation of these cells. Other agents such as lipopolysaccharide and anti-IgM for B cells and concanavalin A for T cells also caused increased IL-4 receptor expression, although to a somewhat ... More
Chemical and enzymatic biotin-labeling of oligodeoxyribonucleotides.
AuthorsKempe T, Sundquist WI, Chow F, Hu SL
JournalNucleic Acids Res
PubMed ID2582352
'Biotin has been converted to 2-(biotinylamido)ethanol and condensed to phosphorylated oligonucleotides in a solid phase synthesis. The 5''-biotinylated oligonucleotides were enzymatically coupled to other DNA fragments by T4 DNA ligase or T4 RNA ligase. The hybridization properties of such biotin-labeled oligonucleotide probes were studied.' ... More
Fluorescence probe studies of the state of tropomyosin in reconstituted muscle thin filaments.
AuthorsIshii Y, Lehrer SS
JournalBiochemistry
PubMed ID3663633
'The monomer fluorescence of N-(1-pyrenyl)maleimide-labeled tropomyosin bound to F-actin (PTm-actin) increases when myosin subfragment 1 (S1) binds to actin and is half complete when only approximately 1 S1 is bound to 7 actin subunits [Ishii, Y., & Lehrer, S. S. (1985) Biochemistry 24, 6631-6638]. Similar studies of the binding of ... More
Development of a dual label fluorescence technique that can be utilized to elucidate the mechanism of action of monoclonal antibody-drug conjugates.
AuthorsStarling JJ, Hinson NA, Marder P, Maciak RS, Laguzza BC, Corvalan JR, Smith W
JournalCancer Res
PubMed ID3048656
'A method is described that allows the simultaneous visualization and relative assessment of both the antibody and drug components of monoclonal antibody-drug conjugates at the target cell membrane. The antibody is detected by a fluorescein-conjugated anti-mouse immunoglobulin serum while the drug is visualized by rhodamine avidin or phycoerythrin-streptavidin binding to ... More
Mode of centriole duplication and distribution.
AuthorsKochanski RS, Borisy GG
JournalJ Cell Biol
PubMed ID2335566
'Centriole stability and distribution during the mammalian cell cycle was studied by microinjecting biotinylated tubulin into early G1 cells and analyzing the pattern of incorporation into centrioles. Cells were extracted and cold treated to depolymerize labile microtubules, allowing the fluorescent microscopic visualization of the stable centrioles. The ability to detect ... More
Nuk controls pathfinding of commissural axons in the mammalian central nervous system.
AuthorsHenkemeyer M, Orioli D, Henderson JT, Saxton TM, Roder J, Pawson T, Klein R
JournalCell
PubMed ID8689685
'Eph family receptor tyrosine kinases have been proposed to control axon guidance and fasciculation. To address the biological functions of the Eph family member Nuk, two mutations in the mouse germline have been generated: a protein null allele (Nuk1) and an allele that encodes a Nuk-beta gal fusion receptor lacking ... More
Enzymatic synthesis of biotin-labeled polynucleotides: novel nucleic acid affinity probes.
AuthorsLanger PR, Waldrop AA, Ward DC
JournalProc Natl Acad Sci U S A
PubMed ID6273878
'Analogs of dUTP and UTP that contain a biotin molecule covalently bound to the C-5 position of the pyrimidine ring through an allylamine linker arm have been synthesized. These biotin-labeled nucleotides are efficient substrates for a variety of DNA and RNA polymerases in vitro. Polynucleotides containing low levels of biotin ... More
A reagent for covalently attaching biotin to proteins via a cleavable connector arm.
AuthorsMouton CA, Pang D, Natraj CV, Shafer JA
JournalArch Biochem Biophys
PubMed ID6756312
Tumor-associated ganglio-N-triosylceramide. Target for antibody-dependent, avidin-mediated drug killing of tumor cells.
AuthorsUrdal DL, Hakomori S
JournalJ Biol Chem
PubMed ID6448851
Bifunctional enkephalin analogues for affinity separation purposes.
AuthorsKoman A, Terenius L
JournalFEBS Lett
PubMed ID6252056
Reversed-phase liquid chromatography of biotin-labelled nucleotides. A new class of markers in molecular biology.
AuthorsToenges V, Reichert A
JournalJ Chromatogr
PubMed ID2415543
Inactivation by avidin of biotin-modified bacteriophage.
AuthorsBecker JM, Wilchek M
JournalBiochim Biophys Acta
PubMed ID4553809
The biotin transport system.
AuthorsBayer EA, Wilchek M
JournalMethods Enzymol
PubMed ID333227
Monoclonal antibody to biotin.
AuthorsDakshinamurti K, Rector ES
JournalMethods Enzymol
PubMed ID2388565
New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates.
AuthorsLiu FT, Zinnecker M, Hamaoka T, Katz DH
JournalBiochemistry
PubMed ID84683
The avidin-biotin complex in affinity cytochemistry.
AuthorsBayer EA, Skutelsky E, Wilchek M
JournalMethods Enzymol
PubMed ID440114
Affinity cytochemistry: the localization of lectin and antibody receptors on erythrocytes via the avidin-biotin complex.
AuthorsBayer EA, Wilchek M, Skutelsky E
JournalFEBS Lett
PubMed ID976475
Progress in enzyme immunoassays: production of reagents, experimental design, and interpretation.
AuthorsKurstak E
JournalBull World Health Organ
PubMed ID3910300
Preparation of modified tubulins.
AuthorsHyman A, Drechsel D, Kellogg D, Salser S, Sawin K, Steffen P, Wordeman L, Mitchison T
JournalMethods Enzymol
PubMed ID2034137
Visualization of intracellular trafficking: use of biotinylated ligands in conjunction with avidin-gold colloids.
AuthorsMorris RE, Saelinger CB
JournalJ Histochem Cytochem
PubMed ID6690597
A simple, sensitive method to visualize the binding and internalization of protein ligands by cells in culture is described. A biotinylated toxin was used as ligand, and succinoylated avidin adsorbed onto 5.2 nm gold sols was the electron-dense marker. This method affords direct localization of proteins that are on the ... More
Polynucleotide-histone H1 complexes as probes for blot hybridization.
AuthorsRenz M
JournalEMBO J
PubMed ID6641717
Biotin and/or 125I-labelled histone H1 proteins (21 kd) have been covalently bound to single-stranded DNA. Complexes of equal masses of DNA and modified histone H1 (approximately 1 histone H1 molecule per 70 nucleotides) were used as probes for blot hybridization experiments and found to have hybridization characteristics very similar (or ... More
Amplification of the biotin-avidin immunmofluorescence technique.
AuthorsBerman JW, Basch RS
JournalJ Immunol Methods
PubMed ID7000912
An amplification of the immunofluorescence technique which uses biotinylated antibody and fluoresceinated avidin is described. By introducing a sandwich technique using fluorescein-conjugated goat anti-avidin, a 5-fold enhancement of staining over the conventional immunofluorescence method is achieved, and the brightness is more than twice that achieved with the simple biotin-fluoresceinated avidin ... More
The quantitation of biotinylated compounds by a solid-phase assay using a 125I-labelled biotin derivative.
AuthorsSmith PJ, Warren RM, von Holt C
JournalFEBS Lett
PubMed ID3582656
The biotin analogue biotinylglycyltyrosine has been synthesized and labelled to a specific activity of 2000 Ci/mmol with 125I. This analogue has been used in conjunction with immobilized streptavidin in an assay which detects as little as 1 fmol biotin or biotinylated molecules in solution. The determination of biotinylated insulin in ... More
A simple method for immunoselective cell separation with the avidin-biotin system.
AuthorsWormmeester J, Stiekema F, De Groot K
JournalJ Immunol Methods
PubMed ID6368688
A new method for immunoselective cell separation is described which uses biotinylated antibodies and avidin-coupled sheep erythrocytes for rosette formation, and a density gradient to separate the rosetting from the non-rosetting cells. The utility of the technique is illustrated by separating subpopulations of rabbit, mouse and rat lymphocytes. The method ... More
Efficient methods for attaching non-radioactive labels to the 5' ends of synthetic oligodeoxyribonucleotides.
AuthorsAgrawal S, Christodoulou C, Gait MJ
JournalNucleic Acids Res
PubMed ID3748808
The syntheses are described of two types of linker molecule useful for the specific attachment of non-radioactive labels such as biotin and fluorophores to the 5' terminus of synthetic oligodeoxyribonucleotides. The linkers are designed such that they can be coupled to the oligonucleotide as a final step in solid-phase synthesis ... More
Use of biotin-labeled monoclonal antibodies and avidin-peroxidase conjugates for the determination of epitope specificities in a solid-phase competitive enzyme immunoassay.
AuthorsWagener C, Fenger U, Clark BR, Shively JE
JournalJ Immunol Methods
PubMed ID6200540
The epitope specificities of monoclonal antibodies against carcinoembryonic antigen (CEA) were determined in a solid-phase, biotin-avidin enzyme immunoassay. Constant amounts of biotin-labeled antibodies were incubated with the immobilized antigen in the presence of decreasing amounts of unlabeled antibodies. The biotinylated antibodies bound to the antigen were then reacted with avidin-peroxidase ... More
A cell nanoinjector based on carbon nanotubes.
AuthorsChen X, Kis A, Zettl A, Bertozzi CR,
JournalProc Natl Acad Sci U S A
PubMed ID17485677
Technologies for introducing molecules into living cells are vital for probing the physical properties and biochemical interactions that govern the cell's behavior. Here, we report the development of a nanoscale cell injection system (termed the nanoinjector) that uses carbon nanotubes to deliver cargo into cells. A single multiwalled carbon nanotube ... More
A novel procedure for labeling immunoglobulins by conjugation to oligosaccharide moieties.
AuthorsO'Shannessy DJ, Dobersen MJ, Quarles RH
JournalImmunol Lett
PubMed ID6394473
A novel method is described for the biotinylation of immunoglobulins. The procedure relies on the generation of reactive aldehydes on the carbohydrate moieties of the immunoglobulin by oxidation with sodium periodate and subsequent reaction with biotin hydrazide. The method is simple and specific and results in stable conjugates retaining full ... More
A biotin-avidin enzyme immunoassay for the quantitation of serum ferritin.
AuthorsLee M, Chang J, Carlson D, Burgett M
JournalClin Chim Acta
PubMed ID3886204
A biotin-avidin enzyme immunoassay (EIA) for measurement of ferritin in human serum is described. This procedure involves the use of specific antibody immobilized on beads, biotin-labeled specific antibody and enzyme-labeled avidin. Reproducible results were achieved within 2 h for ferritin in serum in the range of 5-500 ng/ml. Ferritin levels ... More
Rapid production of quasi-stable antibody-phycoerythrin conjugates for use in flow cytometry.
AuthorsDale GL
JournalCytometry
PubMed ID9845444
A simple system is described for producing quasi-stable complexes between biotinylated antibodies and fluorochrome-labeled streptavidin. Optimal complexes were produced by mixing biotinylated antibody and phycoerythrin-streptavidin together at a molar ratio of 1:1 for 2 min, followed by the addition of soluble biotin to halt oligomerization. These complexes, which are stable ... More
Binding affinities to rat brain synaptosomes--synthesis of biotinylated analogues of Substance P.
AuthorsLavielle S, Chassaing G, Beaujouan JC, Torrens Y, Marquet A
JournalInt J Pept Protein Res
PubMed ID6083984
The synthesis of four biotinylated analogues of Substance P is described. The affinities of these analogues and of their complexes with avidin for the 125I-Bolton Hunter Substance P binding sites on rat brain synaptosomes were determined. While these biotinylated peptides complexed to avidin retain a good biological activity on the ... More
Studies towards the development of chemically synthesized non-radioactive biotinylated nucleic acid hybridization probes.
AuthorsAl-Hakim AH, Hull R
JournalNucleic Acids Res
PubMed ID3027670
Non-radioactive nucleic acid hybridization probes have been constructed in which the reporter group is long chain biotin chemically linked to a basic macromolecule (histone H1, cytochrome C or polyethyleneimine). The modified basic macromolecule which carries many biotin residues can, in turn, be covalently linked to nucleic acids (DNA) via the ... More
Transbilayer movement of phosphatidylserine in erythrocytes: inhibition of transport and preferential labeling of a 31,000-dalton protein by sulfhydryl reactive reagents.
AuthorsConnor J, Schroit AJ
JournalBiochemistry
PubMed ID3365366
A series of labeled thiolation reagents were synthesized on the basis of the parent structure pyridyldithioethylamine (PDA). These compounds specifically and reversibly inhibit the active intrabilayer transport of phosphatidylserine (PS) in human red blood cells. The binding of PDA to cells can be quantified since the thiol-disulfide exchange reaction yields ... More
Biotinylation: an alternative to radioiodination for the identification of cell surface antigens in immunoprecipitates.
AuthorsCole SR, Ashman LK, Ey PL
JournalMol Immunol
PubMed ID3309626
A method has been developed in which the conventional radioiodine label is replaced by non-radioactive biotin in studies involving the immunoprecipitation and analysis of cell surface antigens. The labelling reagent, d-biotinyl-N-hydroxysulfosuccinimide ester (NHSS-biotin), reacts preferentially with lysine residues in polypeptides and possibly also with free amino-groups on carbohydrates and lipids. ... More
Specific localization and quantification of biotin transport components in yeast by use of a biotin-conjugated, impermeant, electron-dense label.
AuthorsBayer EA, Skutelsky E, Viswanatha T, Wilchek M
JournalMol Cell Biochem
PubMed ID347252
Two approaches are described for the localization and quantification of biotin transport components in yeast cells. One approach is based on tracing the fate of a radioactive affinity label for the biotin transport system, [14C]biotinyl-p-nitrophenyl ester (pBNP), through various stages of subcellular fractionations. A complementary method involves the use of ... More
Electron microscopic visualization of the SH1 thiol of myosin by the use of an avidin-biotin system.
AuthorsSutoh K, Yamamoto K, Wakabayashi T
JournalJ Mol Biol
PubMed ID6548525
One of the reactive thiols in the myosin head, SH1, was covalently labeled with a biotin derivative, N-iodoacetyl-N'-biotinylhexylenediamine. When 50% of the SH1 thiol was modified with the biotin reagent as judged from measurements of ATPase activities, the biotinylated myosin bound one mole of avidin per mole of myosin at ... More
Characterization of lactoferrin binding to the MAC-T bovine mammary epithelial cell line using a biotin-avidin technique.
AuthorsRejman JJ, Turner JD, Oliver SP
JournalInt J Biochem
PubMed ID8174755
1. A non-radioisotopic method utilizing a biotin-avidin approach was used to characterize lactoferrin binding to the clonal MAC-T bovine mammary epithelial cell line. 2. Binding of lactoferrin to MAC-T cells and isolated membranes was specific and saturable. 3. Unlabeled lactoferrin competed for and displaced biotin-labeled lactoferrin from binding sites on ... 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
Detection of low-density cell-surface molecules using biotinylated fluorescent microspheres.
AuthorsWojchowski DM, Sytkowski AJ
JournalBiochim Biophys Acta
PubMed ID3964705
Biotinylated fluorescent microspheres have been developed as a reagent for studying antigens and receptors expressed at the cell surface. Labeling of antigen or receptor was accomplished by crosslinking biotinylated microspheres through streptavidin to corresponding biotinylated antibodies or ligands. Detection of labeled cells by flow microfluorimetry provided an extremely sensitive means ... 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
Irreversible inhibition of biotin transport in yeast by biotinyl-p-nitrophenyl ester.
AuthorsBecker JM, Wilchek M, Katchalski E
JournalProc Natl Acad Sci U S A
PubMed ID4944635
Biotinyl-p-nitrophenyl ester (BNP), an active-ester derivative of biotin, irreversibly inactivates biotin transport in the yeast Saccharomyces cerevisiae. Transport inactivation is progressive with time and occurs at concentrations of the ester as low as 10(-7) M. In the presence of sodium azide, a reagent known to block biotin accumulation in yeast, ... More
Measurement of association constants in ELISA. Reactions between solid-phase antibody and fluid-phase biotinylated antigen.
AuthorsMurray JS, Brown JC
JournalJ Immunol Methods
PubMed ID2319139
An ELISA procedure which utilized an avidin-peroxidase amplification reaction to detect and quantify the amount of fluid-phase biotinylated antigen bound to solid-phase antibody was developed to determine antibody association constants. The methodology required is uncomplicated, avoids the use of radioisotopes, and is theoretically amenable for use with any protein which ... More
Transblot studies with biotin-labeled proteins: electrophoretic mobilities and detection limits.
AuthorsNeumaier M, Fenger U, Wagener C
JournalAnal Biochem
PubMed ID2426988
Molecular weight standard proteins, mouse IgG as well as several antigens cross-reacting with the carcinoembryonic antigen (CEA), were biotin labeled, submitted to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and transferred to nitrocellulose. The bound proteins were revealed by the use of avidin-peroxidase conjugates and a suitable substrate. The ratio of N-hydrosuccinimido ... More
Liposome immunoassay by long-lived fluorescence detection.
AuthorsOkabayashi Y, Ikeuchi I
JournalAnalyst
PubMed ID9764512
Immunoassay by fluorescence energy transfer from a europium chelate in liposome to allophycocyanin (APC) was demonstrated. Streptavidin or antibody to biotin was bonded to the liposome containing the europium chelate of 2-naphthoyltrifluoroacetone in the bilayer. When the biotin bonded to APC (APC-BT) was added to the prepared liposomes and the ... More
Site-dependent excited-state dynamics of a fluorescent probe bound to avidin and streptavidin.
AuthorsFürstenberg A, Kel O, Gradinaru J, Ward TR, Emery D, Bollot G, Mareda J, Vauthey E,
JournalChemphyschem
PubMed ID19565577
The excited-state dynamics of biotin-spacer-Lucifer-Yellow (LY) constructs bound to avidin (Avi) and streptavidin (Sav) was investigated using femtosecond spectroscopy. Two different locations in the proteins, identified by molecular dynamics simulations of Sav, namely the entrance of the binding pocket and the protein surface, were probed by varying the length of ... More
Biotinylation reagents for the study of cell surface proteins.
AuthorsElia G,
JournalProteomics
PubMed ID18763706
The extraordinarily stable, non-covalent interaction between avidin and biotin is one of the most commonly exploited tools in chemistry and biology. Methods for derivatization with biotin of a variety of molecules (in particular, proteins) have been introduced, in order to allow their efficient recovery, immobilization and detection with avidin-based reagents. ... More
Identification of distinct domains in CD40 involved in B7-1 induction or growth inhibition.
AuthorsGoldstein MD, Watts TH
JournalJ Immunol
PubMed ID8816387
Binding of CD40 ligand to CD40 on a murine B lymphoma M12 induces B7-1 and inhibits cell growth. We have used M12 lymphomas transfected with wild-type and mutant human CD40 molecules to identify regions of the CD40 cytoplasmic tail involved in these signal transduction events. We find that threonine residues ... More
Properties of the kinetochore in vitro. I. Microtubule nucleation and tubulin binding.
AuthorsMitchison TJ, Kirschner MW
JournalJ Cell Biol
PubMed ID4030893
We have isolated chromosomes from Chinese hamster ovary cells arrested in mitosis with vinblastine and examined the interactions of their kinetochores with purified tubulin in vitro. The kinetochores nucleate microtubule (MT) growth with complex kinetics. After an initial lag phase, MTs are continuously nucleated with both plus and minus ends ... More
Enhancement of immune cellular agglutination by use of an avidin-biotin system.
AuthorsCostello SM, Felix RT, Giese RW
JournalClin Chem
PubMed ID572747
Classically, one can increase the titer of an agglutinating or first antibody with an antiglobulin or second antibody. We have used an avidin-biotin system in place of antiglobulin to similarly extend the agglutination by an initial anticellular antibody. Erythrocytes were agglutinated by adding in succession, caproylamidobiotin-antibody, avidin, and extender (caproylamidobiotin-macromolecule). ... More
A sensitive enzyme assay for biotin, avidin, and streptavidin.
AuthorsBayer EA, Ben-Hur H, Wilchek M
JournalAnal Biochem
PubMed ID3706735
Reciprocal enzyme assays are described for the vitamin biotin and for the biotin-binding proteins avidin and streptavidin. The assays are based on the following steps: (a) biotinylated bovine serum albumin is adsorbed onto microtiter plates; (b) streptavidin (or avidin) is bound to the biotin-coated plates; (c) biotinylated enzyme (in this ... More
Highly sensitive immunoassays based on use of liposomes without complement.
AuthorsLitchfield WJ, Freytag JW, Adamich M
JournalClin Chem
PubMed ID6467563
We describe a novel liposome-based immunoassay in which covalently linked hapten-cytolysin conjugates are used instead of complement and surface-immobilized immunoreagents. Stable, unilamellar liposomes containing entrapped alkaline phosphatase as a marker enzyme were prepared by dialysis of octyl glucoside from suspensions of cholesterol and egg yolk lecithin. The resulting vesicles could ... More
Biotinyl-estradiol derivatives in enzyme immunoassays: structural requirements for optimal antibody binding.
AuthorsTiefenauer LX, Andres RY
JournalJ Steroid Biochem
PubMed ID2194074
The use of the avidin/biotin complex in immunoassays is well documented. No comprehensive studies, however, are available on the structural requirements of the linkage between biotin and small molecules to get an optimal antigen-antibody interaction. We have synthesized seven different biotinylated estradiol derivatives. They were evaluated in an antibody- and ... More
Labeling of platelet surface glycoproteins with biotin derivatives.
AuthorsFabris F, Cordiano I, Mazzuccato M, Pradella P, Randi ML, Girolami A
JournalThromb Res
PubMed ID1329258
We describe a non-radioactive method for the labeling of platelet surface proteins, consisting of platelet protein biotinylation by means of N-hydroxysuccinimido-biotin (NHS-B) and biotin-hydrazide (H-B); NHS-B labels proteins amino residues while H-B binds to periodate-modified sialoglycoproteins. Washed platelets were biotinylated and protein bands were detected after SDS-electrophoresis and western-blot using ... More
Properties of the kinetochore in vitro. II. Microtubule capture and ATP-dependent translocation.
AuthorsMitchison TJ, Kirschner MW
JournalJ Cell Biol
PubMed ID4030894
We have studied the interaction of preformed microtubules (MTs) with the kinetochores of isolated chromosomes. This reaction, which we call MT capture, results in MTs becoming tightly bound to the kinetochore, with their ends capped against depolymerization. These observations, combined with MT dynamic instability, suggest a model for spindle morphogenesis. ... More
Biotin-labeled antigen sandwich enzyme-linked immunosorbent assay (BLA-S-ELISA) for the detection of Japanese encephalitis antibody in human and a variety of animal sera.
AuthorsChang HC, Takashima I, Arikawa J, Hashimoto N
JournalJ Immunol Methods
PubMed ID6381601
A biotin-labeled antigen (BLA) was adapted to a sandwich enzyme-linked immunosorbent assay (S-ELISA) for detection of Japanese encephalitis (JE) antibody in a variety of animal sera. JE antigen was fixed on the wells of a microplate and became bound to the specific antibody which could react with a peroxidase-labeled avidin ... More
Cyclic-AMP modulates downstream events in CD40-mediated signal transduction, but inhibition of protein kinase A has no direct effect on CD40 signaling.
AuthorsGoldstein MD, Cochrane A, Watts TH
JournalJ Immunol
PubMed ID9550383
The role of cAMP/protein kinase A (PKA) in CD40 signal transduction is controversial, with evidence both for and against its importance. In this study we have used a tetracycline-repressible expression system to reversibly express a dominant-negative form of the PKA regulatory subunit type I (PKA-R(G324D)) in a B lymphoma line, ... More
Strand-specific attachment of avidin-spheres to double-stranded poliovirus RNA.
AuthorsRichards OC, Ehrenfeld E, Manning J
JournalProc Natl Acad Sci U S A
PubMed ID218216
Poliovirus-specific double-stranded RNA molecules containing covalently attached protein were coupled with a biotin ester through the protein moiety. Subsequent interaction of the RNA-biotin with avidin attached to electronopaque plastic spheres led to the formation of complexes that were easily visualized in the electron microscope. Avidinspheres were associated only with one ... More
Binding of a curarimimetic toxin from cobra venom to the nicotinic acetylcholine receptor. Interactions of six biotinyltoxin derivatives with receptor and avidin.
AuthorsLobel P, Kao PN, Birken S, Karlin A
JournalJ Biol Chem
PubMed ID3161881
We have reacted N-hydroxysuccinimidyl biotin with the principal curarimimetic toxin in Naja naja siamensis venom, biotinylating each of the five lysine residues and the N-terminal isoleucine. The six monobiotinyl-toxins were isolated by ion-exchange chromatography, and the residue modified in each was identified by peptide mapping and amino acid analysis. We ... More