Many of the biotin labeling kits from pierce use an nhsbiotin reagent without the chromogenic fluorophore. Biotin quantitation with quanttag kit from vector laboratories. Amplite colorimetric biotin quantitation kit aat bioquest. Colorimetric biotin assay kit mak171 technical bulletin. Ez biotin quantitation kit from pierce biocompare product. Featured products include cellsurface protein biotinylation and purification kits, antibody labeling and. The pierce biotin quantitation kit contains a premix of haba and avidin and a biotinylated horseradish peroxidase hrp positive control.
Dilutions must be accounted in the calculation step. Pierce bca protein assay kit brigham young university. The endotoxin concentration in a sample is measured using the pierce lal chromogenic endotoxin quantitation kit via a chromogenic signal generated in the presence of endotoxins. The amount of biotin present can be calculated directly from the decreased absorbance at 500nm. Accuorange protein quantitation kit is a highly sensitive fluorescencebased assay for quantitating purified protein or antibody samples in 96well format. Thermo scientific pierce biotin quantitation kit ez. For proteins of lower molecular weight or multiple biotin labels, less protein can be used.
The thermo scientific pierce biotin quantitation kit contains a premix of haba and avidin and a biotinylated horseradish peroxidase hrp positive control. Desthiobiotin is a singlering, sulfurfree analog of biotin that binds to. Samples can be measured on a microplate absorbance reader at 405nm. If the pierce biotin quantitation kit will be performed to determine the level of biotin incorporation, the protein first must be desalted or dialyzed to remove nonreacted biotin. This data should not be used to interpret actual results.
Proceed to the calculation of moles of biotin per mole of protein section. This colorimetric biotin quantitation kit provides a convenient method for estimating the molar ratio of biotin to protein in biotinprotein conjugates or for quantitating biotin concentration in a solution. Our kit is more accurate than the other suppliers fluorescent biotin assay brand i. The thermo scientific pierce lal chromogenic endotoxin quantitation kit measures the amount of endotoxin in a protein, peptide or antibody sample using the limulus amebocyte lysate lal assay. Thermo scientific haba 4hydroxyazobenzene2carboxylic.
Quantification of biotin concentration in biohrnfs. Pdf ingel detection of biotinprotein conjugates with a. Fluorescence measurement was performed on spectramax gemini xs fluorometer molecular devices with a 485538 nm filter set and 530 nm cutoff. Biotin quantitation kit colorimetric k811 biovision, inc. The thermo scientific fluorescence biotin quantitation kit requires only 10. Free biotin present in solution with this avidinhaba complex will displace the haba dye and cause the absorbance to decrease. When haba is bound to streptavidin, addition of free biotin or biotinylated biomolecules results in its displacement in the reaction. Thermo scientific pierce biotin quantitation kit includes aliquots of haba reagent. Pierce lal chromogenic endotoxin quantitation kit from thermo fisher scientific. Rolling circle transcriptionamplified hierarchically.
Protein analysis, modification and interaction biotinylation kits. Free biotin will displace the haba dye and cause the absorbance to decrease. Thermo scientific pierce protein assay technical handbook nugi. The quanttag biotin kit bdk2000 is designed to determine the amount of free biotin in solution or the number of biotins attached to nucleic acids, proteins, or other macromolecules. Ample materials are provided to perform 25 assays in a cuvette format or 120 assays in a 96well format. The biotin concentration was measured using a piercetm biotin quantitation kit. The haba dye 4 hydroxyazobenzene2carboxylic acid binds to avidin to produce a yelloworange colored complex which absorbs at 500nm. Biotinylation yields were evaluated using either the pierce biotin quantitation kit following the manufacturers protocol thermo fisher scientific or by an sdspage assay 66. Chemiluminescent transcription factor assay kits 29. By contrast, the traditional colorimetric method requires 100 l of sample, and another leading suppliers fluorescent assay kit requires 50 l of sample.
Urine solutions for the biotin doseresponse experiment were prepared by pooling two nonpregnant patient urine samples, resulting in a urine matrix with an hcg concentration free biotin present in solution with this avidinhaba complex will displace the haba dye and cause the absorbance to decrease. Assessment of biotin interference with qualitative point. Biotin quantitation kit, quanttag vector laboratories, inc. Biotin quantitation reagents and kits thermo scientific pierce biotin quantitation kit haba dye reagent and biotinylated protein standard for convenient colorimetric determination of biotinylation levels in antibodies and other proteins.
The convenient assay can be performed either in a cuvette or in a microplate. If the thermo scientific pierce biotin quantitation kit haba assay. Recombinant antigen tpp47biotin for treponema pallidum. Using a chromogenic biotin reagent offers many advantagesyou can use simple uv quantitation to determine biotin incorporation with no need for the haba assay, an additional assay required to determine the extent of biotin incorporation. It isnt always easy to find a trusted source of lab equipment and chemicals like this this pierce lal chromogenic endotoxin quantitation kit. Haba 24hydroxyazobenzene benzoic acid and avidin are the basis of the kit. Thermo scientific pierce lal chromogenic endotoxin. In food, the major part of biotin is covalently bound to protein, leaving only a minor fraction freely. If the drug requires sulfotag conjugation, label the drug following the protocol outlined in the msd. Biovision biotin quantitation kit colorimetric, k811100. Thermo scientific haba 4hydroxyazobenzene2carboxylic acid. Thermo scientific pierce biotin quantitation kit available on gsava contract for federal government customers only. The habaavidin premix is supplied in convenient noweigh microtube packaging, which eliminates the difficulties as sociated with weighing small quantities of reagent.
The assay employs haba 4hydroxyazobenzene2carboxylic acid, a reagent that shows dramatic spectral. This kit can be used to determine accurately the labeling efficiency of biotinlabeled molecules. For more information, or to download product instructions, visit. What is the difference between the trilink biotin labeling. The thermo scientific pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Quicktiter aav quantitation kit provides an efficient system for rapid quantitation of aav titer. The habaavidin premix is supplied in convenient thermo scientific noweigh microtube packaging, which eliminates the difficulties associated with weighing small quantities of reagent. Dec 05, 2011 the quanttag biotin kit bdk2000 from vector laboratories is designed to determine the amount of free biotin in solution or the number of biotins attached to nucleic acids, proteins, or. Current versions of product instructions are available at. Biotin quantitation kit, quanttag vector laboratories. Sep 05, 2014 the biotin content of biotinylated peptide mixtures in samples for ms analysis was determined using fluorescent biotin quantitation kit thermo scientific pierce with a synergy mx microplate reader biotek measuring fluorescent excitationemission at 495520, according to the manufacturers instructions. Introduction biotin vitamin h is present in bacterial, funghi, plants and animals. Example labeling and application data goat antimouse igg product no.
This immundiagnostik assay is intended for the quantitative determination of biotin in serum, plasma, urine and milk. This colorimetric biotin quantitation kit provides a convenient method for estimating the molar ratio of biotin to protein in biotin protein conjugates or for quantitating biotin concentration in a solution. The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. The pulldown assay is an in vitro method used to determine physical interaction between two or more. Assessment of biotin interference with qualitative pointof. The degree of biotin incorporation can be measured with commercial assays such as a haba assay, pierce ez biotin quantitation kit or invitrogen fluoreporter biotin quantitation assay kit. The ez biotin quantitation kit from pierce biotechnology is used to measure the biotin levels in protein samples and thereby measure the molar ratio of biotin to protein in the sample. Biotin quantitation reagents and kits fisher scientific. Titration curve by a double antigen the titer has been suggested in reference to an in. The quanttag biotin kit bdk2000 is designed to determine the amount of free biotin in solution or the number of biotins attached to nucleic acids, proteins. Pierce lal chromogenic endotoxin quantitation kit 88282 from.
This kit can be used to determine accurately the labeling efficiency of biotin labeled molecules. Accuorange protein quantitation kit is a highly sensitive fluorescencebased assay for quantifying purified protein samples in 96well format. Pdf biotin switch assays for quantitation of reversible. Thermo scientific pierce biotin quantitation kit ez biotin. The thermo scientific pierce biotin quantitation kit contains a premix of. Direct detection of biotinylated proteins by mass spectrometry. The labeling rate of the protein with biotin is not only dependend from the reaction stoichiometry, but also from the concentration of the reaction partner.
Fluoreporter biotin quantitation assay kit for biotinylated proteins f30751 figure 1. Pierce lal chromogenic endotoxin quantitation kit from thermo fisher scientific supplier page view company product page pricing info. Introduction the thermo scientific pierce lal chromogenic endotoxin quantitation kit is an efficient, quantitative endpoint assay for the detection of gramnegative bacterial endotoxins. The samples were mixed with a freshly prepared working solution according to the manufacturers protocols, containing dylight reporter working reagent in pbs 2. Abcam s biotin quantitation kit colorimetric ab185441 is based on the differential binding of streptavidin to biotin and a dye, 24hydroxyazobenzene benzoic acid haba.
Procedure for quantitation of moles of biotin per mole of protein microplate format. Determine the molar ratio of biotin incorporated into a. The haba biotin quantitation kit provides a convenient method for estimating the molar ratio of biotin to protein in biotinprotein conjugates or for quantitating biotin concentration in a solution. In the microplate assay format suggested in the ez biotin quantitation kit pierce, the decrease in absorbance for a regular antibody mw 150 kda, at 1 mg. Fluorescence biotin quantitation kit thermo fisher scientific. Bridging immunogenicity assays guidelines for assay development.
The thermo scientific pierce biotinylated protein interaction pulldown kit contains the necessary components to. Thermo scientific pierce biotin quantitation kit includes aliquots of haba reagent and biotinylated protein standard for convenient colorimetric determination of biotinylation levels in labeled antibodies and other proteins. The thermo scientific ez biotin quantitation kit contains premeasured doses of the avidinhaba mixture and biotinylated horseradish peroxidase positive control to simplify reagent preparation and minimize the amount of waste generated. Each kit provides sufficient quantities to perform up to 20 tests for unpurified aav2 or aavdj samples generated from the aav helper free system. The assay utilizes haba 24hydroxyphenylazobenzoic acid, a reagent that shows dramatic spectral changes when bound to avidin. Invitrogen fluoreporter biotin quantitation assay kit. Determine the molar ratio of biotin incorporated into a protein using the habaavidin method. Biotin quantitation kit colorimetric k811 biovision. The thermo scientific pierce ez biotin quantitation kit contains premeasured doses of the avidin. Free amino acids, peptides and low molecular weight proteins do not. Thermo scientific pierce immunostain enhancer 14 thermo scientific dapi stain 15 thermo scientific hoechst 33342 stain 1617 thermo scientific dylight conjugates 1819 licor odyssey instrument products 2021 fluorescence biotin quantitation kit 2223 fluorescent protein gel stains 24. Pierce lal chromogenic endotoxin quantitation kit 88282.
The following figures demonstrate typical results with the gfp fluorometric quantitation kit. Biotinylated protein or macromolecule, free of nonconjugated biotinylation. If the molecule being conjugated to avidin is galactosidase or other free thiol containing. If the level of biotin incorporation will be determined using the pierce biotin quantitation kit haba assay. Urine specimens were stored at 28 c, protected from light, for no longer than 7 days prior to the measurement of biotin and creatinine concentration roche crej2, c701. Pierce biotin quantitation kit thermo fisher scientific. The fluorescence biotin quantitation kit is requires only 10 l of sample. When haba is bound to streptavidin, addition of free biotin or biotinylated biomolecules results in its displacement in.
912 1586 1319 269 513 474 795 670 812 1254 986 587 670 36 319 181 494 1161 33 374 1497 952 913 1295 1352 295 754 947 1168 1437 245 99 794 628 697 1198 1022 601 1002 798 1424 338 542 55 132 594 1261 665 1203 122