AuthorsBardelmeijer HA, Lingeman H, de Ruiter C, Underberg WJ
JournalJ Chromatogr A
PubMed ID9646486
'In recent years capillary electrophoresis (CE) has been developed into a versatile separation technique, next to gas and liquid chromatography (LC), well suited for the determination of a wide variety of e.g., pharmaceutical, biomedical and environmental samples. The main advantages of CE over chromatographic separation techniques are its simplicity and ... More
Focusing of nitric oxide mediated nitrosation and oxidative nitrosylation as a consequence of reaction with superoxide.
AuthorsEspey MG, Thomas DD, Miranda KM, Wink DA
JournalProc Natl Acad Sci U S A
PubMed ID12177414
'The impact of nitric oxide (NO) synthesis on different biological cascades can rapidly change dependent on the rate of NO formation and composition of the surrounding milieu. With this perspective, we used diaminonaphthalene (DAN) and diaminofluorescein (DAF) to examine the nitrosative chemistry derived from NO and superoxide (O2-) simultaneously generated ... More
Mycothiol biosynthesis and metabolism. Cellular levels of potential intermediates in the biosynthesis and degradation of mycothiol in mycobacterium smegmatis.
AuthorsAnderberg SJ, Newton GL, Fahey RC
JournalJ Biol Chem
PubMed ID9804803
'Mycothiol (MSH; 1-D-myo-inosityl-2-(N-acetyl-L-cysteinyl)amido-2-deoxy-alpha-D- glucop yranoside (AcCys-GlcN-Ins)) is a novel thiol produced at millimolar levels by mycobacteria and other actinomycetes that do not make glutathione. We developed methods to determine the major components of MSH (AcCys, Cys-GlcN, AcCys-GlcN, Cys-GlcN-Ins, GlcN-Ins) in cell extracts. Mycobacterium smegmatis was shown to produce measurable levels ... More
Proteolytic activities of human fibroblast collagenase: hydrolysis of a broad range of substrates at a single active site.
AuthorsFields GB, Netzel-Arnett SJ, Windsor LJ, Engler JA, Birkedal-Hansen H, Van Wart HE
JournalBiochemistry
PubMed ID2168739
'The action of human fibroblast collagenase (HFC) on six substrates of markedly different size, sequence, and conformation, including rat type I collagen, rat alpha 1(I) gelatin, beta-casein, and the three synthetic oligopeptides Gly-Pro-Gln-Gly-Ile-Ala-Gly-Gln, Asp-Val-Ala-Gln-Phe-Val-Leu-Thr-Pro-Gly, and Pro-Val-Gln-Pro-Ile-Gly-Pro-Gln, has been examined. The first peptide is a model for the collagenase cleavage site ... More
Analytical methods for the characterization of cationic lipid-nucleic acid complexes.
AuthorsFerrari ME, Nguyen CM, Zelphati O, Tsai Y, Felgner PL
JournalHum Gene Ther
PubMed ID9508052
'Five analytical assays are described that provide a platform for systematically evaluating the effect of formulation variables on the physical properties of cationic lipid-DNA complexes (lipoplexes). The assays are for (i) lipid recovery, (ii) total DNA, (iii) free DNA, (iv) nuclease sensitivity, and (v) physical stability by filtration. Lipid recovery ... More
Human immunodeficiency virus, type 1 protease substrate specificity is limited by interactions between substrate amino acids bound in adjacent enzyme subsites.
AuthorsRidky TW, Cameron CE, Cameron J, Leis J, Copeland T, Wlodawer A, Weber IT, Harrison RW
JournalJ Biol Chem
PubMed ID8617736
'The specificity of the retroviral protease is determined by the ability of substrate amino acid side chains to bind into eight individual subsites within the enzyme. Although the subsites are able to act somewhat independently in selection of amino acid side chains that fit into each pocket, significant interactions exist ... More
Chiral separation of fluorescamine-labeled amino acids using microfabricated capillary electrophoresis devices for extraterrestrial exploration.
AuthorsSkelley AM, Mathies RA
JournalJ Chromatogr A
PubMed ID14735988
'Chiral separations of fluorescamine-labeled amino acids are characterized and optimized on a microfabricated capillary electrophoresis (CE) device. A standard mixture of acidic and neutral amino acids is labeled with fluorescamine in less than 5 min and the hydroxypropyl-beta-cyclodextrin (HPbetaCD) concentration, temperature, and pH are optimized (15 mM HPbetaCD, 6 degrees ... More
Purification, characterization, and localization of follipsin, a novel serine proteinase from the fluid of porcine ovarian follicles.
AuthorsHamabata T, Okimura H, Yokoyama N, Takahashi T, Takahashi K
JournalJ Biol Chem
PubMed ID8027045
'Follipsin, an enzyme that accumulates in the follicular fluid of porcine ovaries during follicular maturation, was purified to apparent homogeneity. The purified enzyme consists of two different polypeptide chains having M(r) = 45,000 and 32,000 each, associated covalently. The enzyme activity was strongly inhibited by diisopropyl fluorophosphate, benzamidine, leupeptin, and ... More
A comparison of fluorescence versus chemiluminescence detection for analysis of the fluorescamine derivative of histamine by HPLC.
AuthorsWalters DL, James JE, Vest FB, Karnes HT
JournalBiomed Chromatogr
PubMed ID7841763
'Fluorescence and chemiluminescence detection were compared for HPLC analysis of the fluorescamine derivative of histamine. The kinetic behaviour of the chemiluminescent response for the derivative was characterized in a static system. An HPLC method was optimized for the derivative using fluorescence detection. Fluorescence detection was linear over the range of ... More
Detection of transformed cells using a fluorescent probe: the molecular basis for the differential reaction of fluorescamine with normal and transformed cells.
AuthorsParry G, Blenis J, Hawkes SP
JournalCytometry
PubMed ID6291884
'Normal and transformed fibroblasts can be discriminated by a flow cytometry assay on the basis of their differential reaction with fluorescamine. The cause of altered reactivity of transformed cells with this fluorescent probe has been investigated by a detailed analysis of its reaction with chicken embryo fibroblasts transformed by a ... More
Fluorimetric determination of procaine in pharmaceutical preparations based on its reaction with fluorescamine.
AuthorsSegura Carretero A, Cruces-Blanco C, Fernández Peinado S, El Bergmi R, Fernández Gutiérrez A
JournalJ Pharm Biomed Anal
PubMed ID10703964
'A simple spectrofluorimetric analysis of a local anaesthetic named procaine using a specific labelling reagent for primary amino groups, has been developed. Because procaine shows very weak native fluorescence, the technique of spectrofluorimetry has been very much limited for its determination. A detail study of the variables affecting the derivatisation ... More
Influences of metal ions on the reaction of amino and imino acids with fluorogenic reagents.
AuthorsMiyano H, Toyo'oka T, Imai K, Nakajima T
JournalAnal Biochem
PubMed ID3936376
'When a twofold excess of metal ions (Cu2+, Ni2+, Zn2+, Mg2+, Ca2+, Fe3+, and Al3+) was added to a reaction medium of amino acids (1-50 microM each) with the fluorogenic reagents NBD-F (4-fluoro-7-nitrobenzo-2-oxa-1,3-diazole), DNS-Cl (5-N,N-dimethylaminonaphthalenesulfonyl chloride), OPA (o-phthalaldehyde), and fluorescamine (4-phenylspiro[furan-2(3H),1''-phthalan]-3,3''-dione), a reduction of derivatization was observed. The effect was ... More
High-performance liquid chromatographic determination of peptides in biological fluids by automated pre-column fluorescence derivatization with fluorescamine.
AuthorsBoppana VK, Miller-Stein C, Politowski JF, Rhodes GR
JournalJ Chromatogr
PubMed ID1939430
'Peptides containing a free alpha- or epsilon-amino group react with fluorescamine under mild alkaline conditions to generate a highly fluorescent but unstable reaction product and, consequently, practical high-performance liquid chromatographic (HPLC) approaches to analysis have typically involved the use of postcolumn derivatization. An automated precolumn approach is reported in which ... More
Homocysteine redox receptor and regulation of extracellular matrix components in vascular cells.
AuthorsTyagi SC
JournalAm J Physiol
PubMed ID9486129
'Dynamic changes in the reduction-oxidation (redox) state of the tissue lead to the pathophysiological condition. Reduced homocysteine causes dysfunctions in endothelium. The proliferation of smooth muscle cells may lead to occlusive vascular disease, ischemia, and heart failure, but whether fibrosis and hypertension are a consequence of smooth muscle proliferation is ... More
Reassociation of the pyruvate dehydrogenase complex from Escherichia coli: Kinetic measurements and binding studies by resonance energy transfer.
AuthorsGraupe K, Abusaud M, Karfunkel H, Bisswanger H
JournalBiochemistry
PubMed ID7041975
On the reactions of fluorescamine with chromosomal proteins.
AuthorsBode J
JournalAnal Biochem
PubMed ID517740
Spectrofluorimetric determination of vigabatrin and gabapentin in urine and dosage forms through derivatization with fluorescamine.
AuthorsBelal F, Abdine H, Al-Majed A, Khalil NY
JournalJ Pharm Biomed Anal
PubMed ID11682233
'A stability-indicating, sensitive, simple and selective spectrofluorimetric method was developed for the determination of vigabatrin (VG) and gabapentin (GB). The method is based on the reaction between the two drugs and fluorescamine in borate buffer of pH 8.2 to give highly fluorescent derivatives that are measured at 472 nm using ... More
LC assay for salmon calcitonin in aerosol formulations using fluorescence derivatization and size exclusion chromatography.
AuthorsWindisch V, Karpenko C, Daruwala A
JournalJ Pharm Biomed Anal
PubMed ID1391086
'A highly sensitive LC procedure was developed that utilizes fluorescence derivatization and detection coupled with size exclusion chromatography for the analysis of salmon calcitonin in salmon calcitonin aerosols. The LC procedure uses fluorescamine derivatization to label the primary amino groups of the peptide. The derivatization procedure is completely automated by ... More
Rapid detection of amphetamine in urine by micro thin-layer chromatography and fluorescence.
AuthorsDecker WJ, Thompson JD
JournalClin Toxicol
PubMed ID750161
'A rapid, inexpensive, and simple screening procedure for the detection of amphetamine abuse was developed for use by laboratories without sophisticated equipment. A small volume of extract from a pH-adjusted urine specimen is used to spot a high-resolution micro TLC plate. The developed TLC plate is sprayed with a solution ... More
The use of fluorescamine as a permeant probe to localize phosphatidylethanolamine in intact friend erythroleukaemic cells.
AuthorsRawyler A, Roelofsen B, Op den Kamp JA
JournalBiochim Biophys Acta
PubMed ID6582936
'Intact Friend erythroleukaemic cells (Friend cells) were incubated at 0-4 degrees C with increasing amounts of fluorescamine. Phospholipids were extracted and the amounts of phosphatidylethanolamine and of its fluorescamine derivative were determined. (1). The plasma membrane of intact Friend cells appeared to be permeable to fluorescamine in a concentration-dependent way. ... More
Functional signal peptide reduces bilayer thickness of phosphatidylcholine liposomes.
AuthorsTahara Y, Murata M, Ohnishi S, Fujiyoshi Y, Kikuchi M, Yamamoto Y
JournalBiochemistry
PubMed ID1390661
'To investigate the interaction between a signal peptide and the lipid bilayer, two kinds of peptides, L8-M5 (L8 = MRL8PLAALG, M5 = KVFER) and L14-M5 (L14 = MRL14PLAALG), were examined in membranes composed of dioleoylphosphatidylcholine (DOPC). Peptides L8 and L14 are artificially designed signal sequences, and M5 is the N-terminal ... More
Simultaneous measurement of cytochrome P4501A catalytic activity and total protein concentration with a fluorescence plate reader.
AuthorsKennedy SW, Jones SP
JournalAnal Biochem
PubMed ID7856852
'A method for simultaneous measurement of the cytochrome P4501A-associated enzyme, ethoxyresorufin-O-deethylase (EROD), and total protein concentration in liver microsomes is described. EROD assays were carried out in multiwell plates, and the fluorescent product (resorufin) and total proteins were quantified within the same wells with a fluorescence plate reader. Fluorescamine was ... More
Simple, rapid and sensitive determination of plasma taurine by high-performance liquid chromatography using pre-column derivative formation with fluorescamine.
AuthorsSakai T, Nagasawa T
JournalJ Chromatogr
PubMed ID1500450
'A simple, rapid and sensitive method for the determination of plasma taurine by high-performance liquid chromatography in the isocratic mode has been developed. The deproteinized plasma was treated with fluorescamine. These derivatives were separated on a LiChrospher 100 RP-8 column within 15 min. The detection limit for taurine was 0.2 ... More
Inhibition of membrane-bound succinate dehydrogenase by fluorescamine.
AuthorsJay D, Jay EG, Garcia C
JournalJ Bioenerg Biomembr
PubMed ID8144496
'Fluorescamine rapidly inactivated membrane-bound succinate dehydrogenase. The inhibition of the enzyme by this reagent was prevented by succinate and malonate, suggesting that the group modified by fluorescamine was located at the active site. The modification of the active site sulfhydryl group by 5,5''-dithiobis(2-nitrobenzoic acid) (DTNB) did not alter the inhibitory ... More
Location and purification of enzymes on polyacrylamide gels using dialyzable fluorescent markers.
AuthorsBodhe AM, Vartak HG, Rele MV, Dhamankar VS
JournalAnal Biochem
PubMed ID3314580
'A method for the location of proteins/enzymes by polyacrylamide gel electrophoresis using dialyzable low-molecular-weight fluorescent peptide markers is described. The markers prepared by treating the peptic digest of casein with fluorescamine showed several bands on gel electrophoresis which helped in locating the proteins. The located desired protein could subsequently be ... More
A sensitive fluorometric assay for reducing sugars.
AuthorsChen F, Liu Y, Lu J, Hwang KJ, Lee VH
JournalLife Sci
PubMed ID1740973
'A simple and rapid fluorometric assay for reducing sugars that is sensitive to the nanomolar range has been developed. The assay involves the derivatization of a given sugar with hydrazine at pH 3 to form a hydrazone, which is reacted with fluorescamine following adjustment of pH to first 9.4 and ... More
A sensitive fluorometric assay for the simultaneous estimation of pepsin and pepsinogen in gastric mucosa.
AuthorsFord TF, Hermon-Taylor J, Grant DA
JournalClin Chim Acta
PubMed ID6816487
'An assay for pepsin has been developed based on the fluorometric measurement of trichloroacetic acid-soluble peptides released from casein at pH 5.3. The increase in relative fluorescence was most sensitive in the range 10-50 micrograms pepsin/l and casein hydrolysis was not affected by the addition of up to a 1000-fold ... More
LC determination of aminoglutethimide enantiomers as dansyl and fluorescamine derivatives in tablet formulations.
AuthorsCesur N, Apak TI, Aboul-Enein HY, Ozkirimli S
JournalJ Pharm Biomed Anal
PubMed ID12008127
'Determination of dansyl (AG-DNS) and fluorescamine (AG-F) derivatives of rac-aminoglutethimide in tablet formulation by HPLC has been achieved on a cellulose tris-(3,5-dimethylphenyl carbamate), known as Chiralcel OD and OD-R under normal and reversed phase columns, respectively, using a fluorescence detector (lambda(ex), 360 nm; lambda(em), 530 nm for AG-DNS derivatives; lambda(ex), ... More
A fluorescence-based protein assay for use with a microplate reader.
AuthorsLorenzen A, Kennedy SW
JournalAnal Biochem
PubMed ID8250247
Gel-staining techniques.
AuthorsMerril CR
JournalMethods Enzymol
PubMed ID1690330
A sensitive, fluorimetric analysis of amino sugars.
AuthorsJimenez MH, Weill CE
JournalCarbohydr Res
PubMed ID7060052
Studies on the reaction of fluorescamine with primary amines.
AuthorsDe Bernardo S, Weigele M, Toome V, Manhart K, Leimgruber W, Böhlen P, Stein S, Udenfriend S
JournalArch Biochem Biophys
PubMed ID4859505
Studies on the kinetics of reaction and hydrolysis of fluorescamine.
AuthorsStein S, Böhlen P, Udenfriend S
JournalArch Biochem Biophys
PubMed ID4859504
Fluorometric assay of secondary amino acids.
AuthorsWeigele M, DeBernardo S, Leimgruber W
JournalBiochem Biophys Res Commun
PubMed ID4689053
Fluorescent labeling of the surface proteins of erythrocyte membranes using cycloheptaamylose-fluorescamine complex.
AuthorsNakaya K, Yabuta M, Iinuma F, Kinoshita T, Nakamura Y
JournalBiochem Biophys Res Commun
PubMed ID1201052
Fluorescent thin-layer peptide mapping for protein identification and comparison in the subnanomole range.
AuthorsStephens RE
JournalAnal Biochem
PubMed ID626356
Fluorescent labeling of mitoplast membrane. Effect of oxidative phosphorylation uncouplers.
AuthorsShiuan D, Tu SI
JournalBiochemistry
PubMed ID667023
A rapid and sensitive fluorescence assay for angiotensin-converting enzyme.
AuthorsConroy JM, Lai CY
JournalAnal Biochem
PubMed ID210690
Sensitive fluorescent determination of trypsin-like proteases.
AuthorsBrown F, Freedman ML, Troll W
JournalBiochem Biophys Res Commun
PubMed ID4795362
A rapid fluorimetric test for lysozyme with purified fluorescamine-labelled peptidoglycan.
AuthorsMoser I, Pittner F, Dworsky P
JournalJ Biochem Biophys Methods
PubMed ID3149658
A sensitive and rapid fluorometric lysozyme assay is described. It is based on the hydrolysis of fluorescamine-labelled peptidoglycan from Micrococcus luteus cell walls. Lysozyme levels as low as 0.1 microgram can be detected. ... More
Determination of long-chain base in glycosphingolipids with fluorescamine.
AuthorsKisic A, Rapport MM
JournalJ Lipid Res
PubMed ID4857650
A method is described for determining the long-chain base content of glycosphingolipids after acid hydrolysis, using the new reagent fluorescamine. The reaction is sensitive and can be used to characterize or measure glycosphingolipids in quantities routinely separated by thin-layer chromatography. ... More
Fluorescamine: a reagent for assay of amino acids, peptides, proteins, and primary amines in the picomole range.
Fluorescamine is a new reagent for the detection of primary amines in the picomole range. Its reaction with amines is almost instantaneous at room temperature in aqueous media. The products are highly fluorescent, whereas the reagent and its degradation products are nonfluorescent. Applications are discussed. ... More
O-phthalaldehyde: fluorogenic detection of primary amines in the picomole range. Comparison with fluorescamine and ninhydrin.
AuthorsBenson JR, Hare PE
JournalProc Natl Acad Sci U S A
PubMed ID1054843
O-Phthalaldehyde, in the presence of 2-mercaptoethanol, reacts with primary amines to form highly fluorescent products. Picomole quantities of amino acids, peptides, and proteins can be detected easily. o-Phthalaldehyde is five to ten times more sensitive than fluorescamine and is soluble and stable in aqueous buffers. ... More
A comparison of fluorescamine and o-phthaldialdehyde as effective blocking reagents in protein sequence analyses by the Beckman sequencer.
AuthorsBhown AS, Cornelius TW, Volanakis JE, Bennett JC
JournalAnal Biochem
PubMed ID6614468
Use of o-phthaldialdehyde to chemically reduce the newly generated amino termini responsible for the progressively increasing background during an extended amino acid sequence analysis in a liquid phase sequencer has been described. The results have been compared with Fluram blocking using apomyoglobin and rabbit C-reactive protein as standard and unknown ... More
Derivatization of posttranslationally modified amino acids.
AuthorsTeerlink T
JournalJ Chromatogr B Biomed Appl
PubMed ID7820276
After a brief overview of posttranslational modifications of protein amino acids, the use of various derivatizing reagents for amino acid analysis is discussed. Derivatization and chromatographic separation of hydroxyproline, methylhistidine, and phosphorylated amino acids are discussed in detail to illustrate some of the strategies that can be applied to the ... More
Use of a rapid and highly sensitive fluorescamine-based procedure for the assay of plasma lipoproteins.
AuthorsFunk GM, Hunt CE, Epps DE, Brown PK
JournalJ Lipid Res
PubMed ID3760715
A rapid and sensitive method for determining protein concentrations using fluorescamine has been characterized for use in the analysis of intact lipoproteins. It was shown that there is no interference with the assay due to the presence of lipid-associated turbidity or primary amine content. The assay was shown to be ... More
Application of fluorescamine to the study of protein-DNA interactions.
AuthorsBode J, Willmitzer
JournalNucleic Acids Res
PubMed ID1237881
The reactivity of alpha-amino groups of basic proteins towards fluorescamine is essentially abolished if salt linkages with DNA phosphate groups are formed. This observation prompted the elaboration of a very general assay which allows the determination of binding parameters for the interaction of proteins with DNA and chromatin. Protamines, labeled ... More
Recent advances in capillary electrophoresis of proteins, peptides and amino acids.
AuthorsNovotny MV, Cobb KA, Liu JP
JournalElectrophoresis
PubMed ID2257844
The current status of high-performance capillary electrophoresis as an analytical separation method for proteins, peptides and amino acids is assessed. Recent advances in suppressing the effects of electroosmotic flow and irreversible adsorption of proteins at the capillary wall are reviewed, together with procedures for optimal separations of peptides and amino ... More
Determination of endotoxin using fluorescent probe.
AuthorsYoshida K, Ozawa A, Saifuku K, Namihisa T, Nanbu M
JournalClin Chim Acta
PubMed ID7094342
Bacterial endotoxin induces gel-formation of amebocyte lysate. This gelation test, the limulus test, was first described by Levin and Bang [1]. The limulus test has been widely used for the detection of endotoxin, not only in fundamental bacteriological research but also in clinical diseases [2]. Although the technique of the ... More
Fluorescence studies on aged and young erythrocyte populations.
AuthorsGareau R, Goulet H, Chénard C, Caron C, Brisson GR
JournalCell Mol Biol
PubMed ID2059983
Structural changes in red blood cell (RBC) membrane are investigated by fluorescence techniques. Results obtained with three probes (DPH, 3-PM and fluorescamine) indicate a significant increase in membrane rigidity associated with aging of RBCs. Discrepancies between our observations and published data could arise from utilization of experimental conditions closer to ... More
Efficient analysis of cytochrome P4501A catalytic activity, porphyrins, and total proteins in chicken embryo hepatocyte cultures with a fluorescence plate reader.
AuthorsKennedy SW, Jones SP, Bastien LJ
JournalAnal Biochem
PubMed ID7793639
An efficient method for measuring polychlorinated biphenyl-mediated induction of cytochrome P4501A, using the ethoxyresorufin-O-deethylase (EROD) assay, and total porphyrins in chicken embryo hepatocyte cultures is described. Hepatocytes were cultured in 48-well plates, assays were carried out within the wells, and concentrations of the product of the EROD reaction (resorufin), porphyrins, ... 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
Drosophila mitotic domain boundaries as cell fate boundaries.
AuthorsCambridge SB, Davis RL, Minden JS
JournalScience
PubMed ID9242613
Fate determination in Drosophila embryos is evidenced by the appearance of mitotic domains. To identify fate or fates of cells, individual cells in mitotic domains 2, 8, and 15 were marked and monitored through development. Comparison of the different fates indicated that domain boundaries are cell fate boundaries. Cells were ... More
Quantification of lysophosphatidylethanolamine in the nanomole range.
AuthorsSchmid PC, Pfeiffer DR, Schmid HH
JournalJ Lipid Res
PubMed ID7288296
A microanalytical procedure for the determination of trace amounts of lysophosphatidylethanolamine and other amine-containing lipids is described. The technique involves reaction of lipid extracts with fluorescamine to give fluorescent derivatives of lipids containing a free amino group, separation of the products by thin-layer chromatography, and quantification by fluorescence spectroscopy after ... More
Fluorimetric assay of renin.
AuthorsGalen FX, Devaux C, Grogg P, Menard J, Corvol P
JournalBiochim Biophys Acta
PubMed ID566120
A simple fluorimetric assay was set up to test renin within 2 h. N-acetyltetradecapeptide was synthesized and used as substrate. It was demonstrated that N-acetyl-angiotensin I and Leu-Val-Tyr-Ser were the two peptides obtained after hydrolysis by renin. Fluorescamine reaction reacted with the free NH2 of the tetrapeptide generated to induce ... More
Fluorescamine as a histochemical reagent: demonstration of polypeptide hormone-secreting cells.
AuthorsLarsson LI, Sundler F, Håkanson R
JournalHistochemistry
PubMed ID1102500
Fluorescamine is a useful fluorescence microscopic reagent for the demonstration of certain peptide hormone-secreting cells in formaldehyde-fixed tissues. Among the cells demonstrated are the pituitary GH cells, the gastrin cells, the insulin cells and the thyroid C cells. In the latter cell system degranulation brings about a marked decrease in ... More
Quantitation of sulfamethazine in pork tissue by thin-layer chromatography.
AuthorsUnruh J, Schwartz DP, Barford RA
JournalJ AOAC Int
PubMed ID8471859
Our earlier method to detect and quantitate sulfamethazine (SMZ) in milk at the 10 ppb level was modified to quantitate SMZ in pork tissue. Sulfabromomethazine (SBZ) is added to the tissue as an internal standard. SMZ and SBZ are extracted from the tissue into water as the supernatant of a ... More
Enzymatic fluorometric assay for plasma pyridoxal 5'-phosphate.
AuthorsPoon HC, Schmidt BM
JournalClin Biochem
PubMed ID2040086
An enzymatic fluorometric assay to quantitate plasma pyridoxal 5'-phosphate (PLP) is described. PLP is preincubated for 30 min with purified tyrosine decarboxylase apoenzyme (TDA) in acetate buffer and is then incubated with L-tyrosine for 60 min. The decarboxylated metabolite, tyramine, is extracted into ethyl acetate, air dried, dissolved in borate ... More
Simplified fluorometric assay for sphingosine bases.
AuthorsHiggins TJ
JournalJ Lipid Res
PubMed ID6491534
A simplified procedure for fluorometric determination of sphingosine bases is presented. Nitrogenous bases are reacted with fluorescamine and then extracted into a lower phase solvent, obviating the need to transfer the reaction products prior to quantitation. Consequently, the entire procedure may be carried out in a single set of screw-capped ... More
Postcolumn derivatization of peptides with fluorescamine in capillary electrophoresis.
AuthorsZhu R, Kok WT
JournalJ Chromatogr A
PubMed ID9718696
Fluorescamine is used as a postcolumn derivatization reagent for fluorescence detection detection of peptides after separation by capillary electrophoresis. The problems resulting from the use of an organic solvent have been solved by introducing LiC1O4 and 5% water into the postcolumn derivatization reagent. The reaction rate and detection sensitivity of ... More
Capillary electrophoretic separations of peptides using micelle-forming compounds and cyclodextrins as additives.
AuthorsLiu J, Cobb KA, Novotny M
JournalJ Chromatogr
PubMed ID2077044
The value of electrokinetic capillary chromatography for separating structurally similar model peptides and tryptic digests is demonstrated. The behavior of model peptides in buffer systems containing dodecyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, sodium dodecyl sulfate and two cyclodextrins as additives is described. These additives, under different analytical circumstances, exhibit certain beneficial effects ... More
Fluorometric determination of nanogram quantities of protein in small samples: application to calcium-transport adenosine triphosphatase.
AuthorsBridges MA, McErlane KM, Kwong E, Katz S, Applegarth DA
JournalClin Chim Acta
PubMed ID2941185
A fluorometric micro protein assay based on fluorescamine-labelling of homogeneous proteins in solution has been developed which is capable of accurately quantitating as little as 25 ng protein at a concentration of 1.25 micrograms/ml. This micro assay uses a flow-through HPLC fluorescence detector. Typical micro assays measuring bovine serum albumin ... More
Bilayer distribution of phosphatidylserine and phosphatidylethanolamine in lipid vesicles.
AuthorsRoy MT, Gallardo M, Estelrich J
JournalBioconjug Chem
PubMed ID9404670
The distribution of phosphatidylethanolamine (PE) and phosphatydilserine (PS) in liposomes was studied as a function of aminophospholipid concentration using fluorescamine as labeling reagent. The method is suitable for such determination since, in the assay conditions, fluorescamine does not penetrate the vesicles nor does it disrupt them. The liposomes were obtained ... More
Accumulation of pyrraline-modified albumin in phagocytes due to reduced degradation by lysosomal enzymes.
AuthorsMiyata S, Liu BF, Shoda H, Ohara T, Yamada H, Suzuki K, Kasuga M
JournalJ Biol Chem
PubMed ID9020111
Previous studies suggested that the interaction between proteins modified by advanced glycation end products (AGEs) and cells, such as macrophages, may be involved in diabetic angiopathy. Pyrraline is one of the AGEs and known to be elevated in plasma of diabetic rats and humans, and is present in vascular lesions ... More
Quantitative fluorometric analysis of plant and microbial chitosanases.
AuthorsOsswald WF, McDonald RE, Niedz RP, Shapiro JP, Mayer RT
JournalAnal Biochem
PubMed ID1514694
A quantitative fluorometric assay for chitosanase activity in bacterial and plant tissues was developed. The assay can be conducted with either finely milled preparations of chitosan in suspension or dissolved chitosan; activity is based on measurements of glucosamine (GlcN) or oligomers of GlcN. GlcN is detected fluorometrically after reaction with ... More
A comparison of fluorescamine and naphthalene-2,3-dicarboxaldehyde fluorogenic reagents for microplate-based detection of amino acids.
AuthorsBantan-Polak T, Kassai M, Grant KB
JournalAnal Biochem
PubMed ID11673879
The use of appropriate fluorometric derivatization procedures is of considerable importance for accurate determination of amino acids in biological samples and in metal-assisted peptide hydrolysis reactions. It is especially critical for the relative fluorescence intensities (RFI) of equal amounts of amino acids to be as similar as possible. While fluorescamine ... More
A fluorescamine assay for membrane protein and peptide samples with non-amino-containing lipids.
AuthorsChung LA
JournalAnal Biochem
PubMed ID9177744
A method is described for determining the concentration of membrane proteins and peptides in the presence of non-amino-containing lipids. The assay is quantitative when used for purified proteins and peptides of known sequence and qualitative when sequences are unknown or samples contain contaminating proteins. In this method, proteins and peptides ... More
Evaluation of six fluorescent protein stains for use in flow microfluorometry.
AuthorsFreeman DA, Crissman HA
JournalStain Technol
PubMed ID52920
Flow microfluorometric (FMF) analysis of stained cells has provided protein distribution histograms for large populations of cells. Spectral data and staining protocols were evaluated for six fluorescent protein dyes suggested for staining cells in liquid suspension. The requirements for dyes and/or staining protocol included minimal cell clumping and cell loss, ... More
Multiphoton-excited fluorescence of fluorogen-labeled neurotransmitters.
AuthorsShear JB, Brown EB, Webb WW
JournalAnal Chem
PubMed ID8651483
Fluorescence detection of fluorogen-labeled neurotransmitters is demonstrated using 100 fs pulses from a titanium-sapphire mode-locked laser to achieve molecular excitation by simultaneous absorption of two and three photons of near-IR radiation. Two-photon excitation spectra are determined for the naphthalene-2,3 dicarboxaldehyde derivative of glycine and the fluorescamine derivative of leucine enkephalin, ... More
A whole new way of looking at things: the use of Dark Reader technology to detect fluorophors.
AuthorsSeville M
JournalElectrophoresis
PubMed ID11332748
The Dark Reader optical system (Clare Chemical Research, Denver, CO, USA) uses relatively low intensity broad-band visible blue light in combination with broad-band optical filters to detect fluorescence with a level of sensitivity that often surpasses that of UV transilluminators and can rival that of laser-based scanners. Applications of DR ... More
Rapid transmembrane movement of newly synthesized phosphatidylethanolamine across the inner membrane of Escherichia coli.
AuthorsHuijbregts RP, de Kroon AI, de Kruijff B
JournalJ Biol Chem
PubMed ID9668071
For the first time the transmembrane movement of an endogenously synthesized phospholipid across the inner membrane of E. coli is reported. [14C]phosphatidylethanolamine (PE) was biosynthetically introduced into inner membrane vesicles from the PE-deficient strain AD93, by reconstitution with the enzyme phosphatidylserine (PS) synthetase. Upon addition of wild type cell lysate ... More
Selective degradation of oxidized calmodulin by the 20 S proteasome.
AuthorsFerrington DA, Sun H, Murray KK, Costa J, Williams TD, Bigelow DJ, Squier TC
JournalJ Biol Chem
PubMed ID11010965
We have investigated the mechanisms that target oxidized calmodulin for degradation by the proteasome. After methionine oxidation within calmodulin, rates of degradation by the 20 S proteasome are substantially enhanced. Mass spectrometry was used to identify the time course of the proteolytic fragments released from the proteasome. Oxidized calmodulin is ... More
Stability of poly(L-lysine)-complexed plasmid DNA during mechanical stress and DNase I treatment.
AuthorsCapan Y, Woo BH, Gebrekidan S, Ahmed S, DeLuca PP
JournalPharm Dev Technol
PubMed ID10578502
The aim of this study was to investigate the formation and stability of complexes between plasmid DNA (pDNA) and poly(L-lysine) (PLL). Formation of pDNA/PLL complexes with various ratios was determined by a fluorescence spectrophotometric method using fluorescamine. The effects of sonication, vortexing, and exposure to DNase I on the stability ... More
The sizes of peptides generated from protein by mammalian 26 and 20 S proteasomes. Implications for understanding the degradative mechanism and antigen presentation.
AuthorsKisselev AF, Akopian TN, Woo KM, Goldberg AL
JournalJ Biol Chem
PubMed ID9920878
Knowledge about the sizes of peptides generated by proteasomes during protein degradation is essential to fully understand their degradative mechanisms and the subsequent steps in protein turnover and generation of major histocompatibility complex class I antigenic peptides. We demonstrate here that 26 S and activated 20 S proteasomes from rabbit ... More
A role for oxidative stress in apoptosis: oxidation and externalization of phosphatidylserine is required for macrophage clearance of cells undergoing Fas-mediated apoptosis.
AuthorsKagan VE, Gleiss B, Tyurina YY, Tyurin VA, Elenström-Magnusson C, Liu SX, Serinkan FB, Arroyo A, Chandra J, Orrenius S, Fadeel B
JournalJ Immunol
PubMed ID12077280
Exposure of phosphatidylserine (PS) on the surface of apoptotic cells has been suggested to serve as an important recognition signal for macrophages. In this work we show that triggering of the death receptor Fas on Jurkat cells results in the generation of reactive oxygen species with oxidation and externalization of ... More
The nitric oxide congener nitrite inhibits myeloperoxidase/H2O2/ Cl- -mediated modification of low density lipoprotein.
AuthorsCarr AC, Frei B
JournalJ Biol Chem
PubMed ID11054430
Nitric oxide, a pivotal molecule in vascular homeostasis, is converted under aerobic conditions to nitrite. Recent studies have shown that myeloperoxidase (MPO), an abundant heme protein released by activated leukocytes, can oxidize nitrite (NO(2-)) to a radical species, most likely nitrogen dioxide. Furthermore, hypochlorous acid (HOCl), the major strong oxidant ... More
Application of redox enzymes for probing the antigen-antibody association at monolayer interfaces: development of amperometric immunosensor electrodes.
AuthorsBlonder R, Katz E, Cohen Y, Itzhak N, Riklin A, Willner I
JournalAnal Chem
PubMed ID8797376
Insulation of the electrical contact between a redox protein and an electrode surface upon association of an antibody to an antigen monolayer assembled on the electrode is used to develop immunosensor devices. In one configuration, a mixed monolayer consisting of the N epsilon-(2,4-dinitrophenyl)lysine antigen and ferrocene units acting as electron ... More
Solid-phase clean-up and thin-layer chromatographic detection of veterinary aminoglycosides.
AuthorsMedina MB, Unruh JJ
JournalJ Chromatogr B Biomed Appl
PubMed ID7704199
Chemical methods are needed to confirm the presence of antibiotics detected by microbial inhibition assays in fluids and tissues of farm animals. We have optimized the conditions for the isolation of hygromycin B with a copolymeric bonded solid-phase silica column followed by thin-layer chromatography (TLC) separation and detection of its ... More
Nitric oxide dissociates lipid oxidation from apoptosis and phosphatidylserine externalization during oxidative stress.
AuthorsFabisiak JP, Tyurin VA, Tyurina YY, Sedlov A, Lazo JS, Kagan VE
JournalBiochemistry
PubMed ID10625487
Oxidative stress in biological membranes can regulate various aspects of apoptosis, including phosphatidylserine (PS) externalization. It is not known, however, if the targets for these effects are lipids or proteins. Nitric oxide (NO), a bifunctional modulator of apoptosis, has both antioxidant and prooxidant potential. We report here that the NO ... More
Decreased binding to proteins and cells of polymeric gene delivery vectors surface modified with a multivalent hydrophilic polymer and retargeting through attachment of transferrin.
AuthorsDash PR, Read ML, Fisher KD, Howard KA, Wolfert M, Oupicky D, Subr V, Strohalm J, Ulbrich K, Seymour LW
JournalJ Biol Chem
PubMed ID10660529
Binding of serum proteins to polyelectrolyte gene delivery complexes is thought to be an important factor limiting bloodstream circulation and restricting access to target tissues. Protein binding can also inhibit transfection activity in vitro. In this study a multivalent reactive hydrophilic polymer has been used to inhibit protein binding. This ... More
Role of functional groups of human plasma and luminol in scavenging of NaOCl and neutrophil-derived hypochlorous acid.
AuthorsArnhold J, Hammerschmidt S, Arnold K
JournalBiochim Biophys Acta
PubMed ID1655046
Hypochlorous acid HOCl/OCl- and other oxidants derived from stimulated polymorphonuclear leukocytes are involved in tissue damage during a number of pathological processes. In order to obtain more detailed information on possible reactions of HOCl/OCl- the effects of both NaOCl and PMN-derived hypochlorous acid on functional groups of amino acid solutions ... More
A nonradioactive fluorescent gel-shift assay for the analysis of protein phosphatase and kinase activities toward protein-specific peptide substrates.
AuthorsLutz MP, Pinon DI, Miller LJ
JournalAnal Biochem
PubMed ID7978268
Synthetic peptides are important tools with which to study the activities of protein kinases and phosphatases toward specific substrate sequences which are present within selected regions of a protein. Most existing assays for the phosphorylation or dephosphorylation of such peptides utilize 32P and either affinity chromatography or HPLC separation and ... More
Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages.
During normal tissue remodeling, macrophages remove unwanted cells, including those that have undergone programmed cell death, or apoptosis. This widespread process extends to the deletion of thymocytes (negative selection), in which cells expressing inappropriate Ag receptors undergo apoptosis, and are phagocytosed by thymic macrophages. Although phagocytosis of effete leukocytes by ... More
Development of a vital fluorescent staining method for monitoring bacterial transport in subsurface environments.
Previous bacterial transport studies have utilized fluorophores which have been shown to adversely affect the physiology of stained cells. This research was undertaken to identify alternative fluorescent stains that do not adversely affect the transport or viability of bacteria. Initial work was performed with a groundwater isolate, Comamonas sp. strain ... More