This kit can be used to determine accurately the labeling efficiency of biotinlabeled molecules. Dilutions must be accounted in the calculation step. 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. The pierce mass spec sample prep kit for cultured cells contains all the necessary reagents and enzymes to prepare up to 20 samples 1 million cells. Assessment of biotin interference with qualitative pointof. It consists of labeling cell surface proteins with a biotin reagent before lysing the cells, and isolating these tagged proteins by. Pdf biotin switch assays for quantitation of reversible.
Colorimetric biotin assay kit mak171 technical bulletin. Biotin quantitation kit colorimetric k811 biovision. Many of the biotin labeling kits from pierce use an nhsbiotin reagent without the chromogenic fluorophore. Biotin quantitation with quanttag kit from vector laboratories. The assay employs haba 4hydroxyazobenzene2carboxylic acid, a reagent that shows dramatic. Conjugation of oligosaccharides to probes and surfaces is useful in the development of biochemical assays to assess carbohydrateprotein interactions. L of the habaavidin premix solution from step 2 to the cuvette. Biotin quantitation reagents and kits fisher scientific. The habaavidin premix is supplied in convenient noweigh microtube packaging, which eliminates the difficulties as sociated with weighing small quantities of reagent. The accublue high sensitivity assay is also available without any dna standards 31008, for those that wish to use their own standard. The thermo scientific pierce lal chromogenic endotoxin quantitation kit is an efficient, quantitative endpoint assay for the detection of gramnegative bacterial endotoxins.
Biovision biotin quantitation kit colorimetric, k811100. Bacterial endotoxin catalyzes theactivation of a proenzyme in the modified limulus amebocyte lysate lal. Samples can be measured on a microplate absorbance reader at 405nm. 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. Piercetm bca protein assay kit thermopierce, catalog number. Enzymatic method for determining lamino acids lamino. This kit can be used to determine accurately the labeling efficiency of biotin labeled molecules. The quanttag biotin kit bdk2000 from vector laboratories is designed to determine the amount of free biotin in solution or the number of. Direct streptavidinbinding assay does not accurately quantitate. The thermo scientific biotin 3 end dna labeling kit uses tdt to incorporate biotinylated ribonucleotides onto the 3 end of dna strands. All samples and standards should be run in duplicate.
The bca protein assay kit provides a simple, rapid, detergent tolerant up to 5% procedure for determining the concentration of proteins in solution. Proceed to the calculation of moles of biotin per mole of protein section. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Pdf quantitative recovery of biotinylated proteins from. Biotin quantitation kit colorimetric k811 biovision, inc.
The thermo scientific fluorescence biotin quantitation kit requires only 10. Pdf ingel detection of biotinprotein conjugates with a. Thermo scientific haba 4hydroxyazobenzene2carboxylic. Thermo scientific pierce biotin quantitation kit includes aliquots of haba reagent. 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. Determine the molar ratio of biotin incorporated into a protein using the habaavidin method. Our kit is more accurate than the other suppliers fluorescent biotin assay brand i. The sensolyte biotin quantitation assay kit provides a convenient assay for estimating the molar ratio of biotin to protein in biotinprotein conjugates or for quantifying biotin concentration in a solution. Cell biolabs high sensitivity protein quantitation assay kit utilizes a fluorogenic detection dye to measure primary aliphatic amines peptides or proteins in various samples with much greater sensitivity than the conventional colorimetric protein determination methods. The assay is performed in a 96well microtiter plate format and measured within 15 minutes. The tdt prefers to label singlestranded dna, but can also label doublestranded dna with blunt or 3 overhangs, but at a lower efficiency. 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. The assay utilizes haba 24hydroxyphenylazobenzoic acid, a reagent that shows dramatic spectral changes when bound to avidin.
Collectively, they represent the stateoftheart for colorimetric detection and quantitation of total protein. Ez biotin quantitation kit from pierce biocompare product. By contrast, the traditional colorimetric method requires 100 l of sample, and another leading suppliers fluorescent assay kit requires 50 l of sample. Highly purified distilled and deionized water was used throughout. The assay employs haba 4hydroxyazobenzene2carboxylic acid, a reagent that shows dramatic spectral changes when bound to avidin.
A modified microbradford procedure for elimination of interference from sodium dodecyl sulfate, other detergents and. It isnt always easy to find a trusted source of lab equipment and chemicals like this this pierce lal chromogenic endotoxin quantitation kit. May 24, 2007 the biotin 3 end dna labeling kit from pierce biotechnology incorporates biotinylated ribonucleotides onto the 3 end of dna strands by using terminal deoxynucleotidyl transferase tdt. 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. 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. Sensolyte biotin quantitation kit fluorimetric size. Development of a sensitive, rapid, biotinstreptavidin. Biotin quantitation kit, quanttag vector laboratories, inc.
Pierce offers seven colorimetric assays for detection and quantitation of total protein. An assay for biotin that uses europium coupled to streptavidin and. 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. Biotin vitamin h is an important biomolecule that has been widely used in biotinylation reactions to label antibodies or other proteins of interest. What is the difference between the trilink biotin labeling. The thermo scientific pierce biotin quantitation kit contains a premix of haba and avidin and a biotinylated horseradish peroxidase hrp positive control. In parallel, a set of known biotin concentrations was used to construct a standard. Amplite colorimetric biotin quantitation kit aat bioquest. Thermo scientific pierce haba is 4hydroxyazobenzene2carboxylic acid, a simple reagent that enables spectrophotometric colorimetric estimation of biotinylation levels of labeled proteins and other molecules. The pierce biotin quantitation kit contains a premix of haba and avidin and a biotinylated horseradish peroxidase hrp positive control. Ingel detection of biotinprotein conjugates with a green fluorescent streptavidin probe.
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. Sulfosuccinimidyl6biotinamido hexanoate nhslcbiotin was purchased from pierce chemical rockford, il. Procedure for quantitation of moles of biotin per mole of protein microplate format. The protein quantification kit provides a simple and rapid procedure for determining the concentration of protein in solution.
This labeling strategy has the advantage of localizing the biotin to the 3 end of the probe where it is less likely to interfere with hybridization or sequencespecific binding of proteins. The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. Cell surface protein biotinylation and analysis bioprotocol. Assessment of biotin interference with qualitative point. Thermo scientific pierce lal chromogenic endotoxin. Quicktiter aav quantitation kit provides an efficient system for rapid quantitation of aav titer for both viral supernatant and purified virus. Ample materials are provided to perform 100 assays in cuvette format or 500 assays in a 96well format. Ez biotin quantitation kit was purchased from pierce usa. Pierce bca protein assay kit brigham young university. 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 pdf. Ezlink sulfonhs biotin sulfosuccinimidyl6 biotin amidohexanoate, 25mg. The biotin 3 end dna labeling kit from pierce biotechnology incorporates biotinylated ribonucleotides onto the 3 end of dna strands by using terminal deoxynucleotidyl transferase tdt. Biotin 3 end dna labeling kit from pierce biotechnology. Thermo scientific pierce protein assay technical handbook.
Meridian road rockford, ll 61105 usa 815 9687316 fax number description. Thermo scientific pierce biotin quantitation kit ez biotin. The thermo scientific pierce bca protein assay kit is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. The kit has a detection sensitivity limit of 1 x 109 gcml genome copyml for unpurified aav2 or aavdj supernatants, or 5 x 10 10 gcml for purified aav sample from any serotype, which is sufficient for mid or hightiter aav samples. They are all wellcharacterized, robust assays that provide consistent, reliable results.
The convenient assay can be performed either in a cuvette or in a microplate. Pierce biotechnology po box 117 815 9680747 3747 n. Both nucleic acid and protein quantitation can be performed in single tube or microplate assays. Biotin, a 244 dalton vitamin found in tiny amounts in all living cells, binds with high affinity to avidin, streptavidin and thermo scientific neutravidin biotinbinding protein. Pierce biotin quantitation kit thermo fisher scientific. Thermo scientific pierce biotin quantitation kit ez. 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. Pdf the strong interaction between streptavidin and biotin is one of the most. For proteins of lower molecular weight or multiple biotin labels, less protein can be used. 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. Thermo scientific pierce biotin quantitation kit available on gsava contract for federal government customers only. The habaavidin premix is supplied in convenient thermo scientific noweigh microtube packaging, which eliminates the difficulties associated with weighing small quantities of reagent.
Haba 24hydroxyazobenzene benzoic acid and avidin are the basis of the kit. Since biotin is a relatively small molecule, it can be conjugated to many proteins without significantly altering their biological activity. Protein quantitation kit is used for determining the concentration of protein in solution. Fluoreporter biotin quantitation assay kit for biotinylated proteins f30751 figure 1. Fluorescence biotin quantitation kit thermo fisher scientific. Pierce biotin quantitation kit thermo fisher scientific the thermo scientific pierce biotin quantitation kit contains a premix of. The accublue high sensitivity dsdna quantitation kit offers sensitive and selective detection of purified dsdna samples with a linear detection range from 0. Assay is suitable for measuring protein concentration in the range of 0. 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. Application methods p a g e 1 080409 nucleic acid and protein quantitation methods prepared by.
Table 1 comparison of common dna, rna and protein quantitation assays method detection. 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. 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. 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. The fluorescence biotin quantitation kit is requires only 10 l of sample. Thermo scientific pierce fluorescence biotin quantitation kit requires only ten microliters of sample to accurately measure the biotinylation level of labeled. The activated proenzyme then catalyzes the splitting of p. Piercea protein assay kit bc thermo fisher scientific. The thermo scientific pierce biotin quantitation kit contains a premix of. The thermo scientific pierce mass spec sample prep kit for cultured cells is an easytouse, comprehensive kit for preparation of clean peptide mixtures from cultured cells for ms analysis. 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.
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