Colorimetric biotin assay kit catalog number mak171 storage temperature 20 c technical bulletin product description biotinylation is the process of labelling a molecule with a biotin molecule to take advantage of the strong binding and high specificity of biotin and streptavidin for purification or detection applications. Furthermore, detection of protein biotinylation using stvgfp was easily detectable with 75 pmol poiavitag under nonoptimal uv excitation conditions, comparing favourably with the linear range of a commercial fluorescence biotin quantitation kit pierce, which has a linear detection range of 1060 pmol. Our kit is more accurate than the other suppliers fluorescent biotin assay. The thermo scientific pierce lal chromogenic endotoxin quantitation kit is an efficient, quantitative endpoint assay for the detection of gramnegative bacterial endotoxins.
Pdf ingel detection of biotinprotein conjugates with a. Calculate results directly from absorbance values based on extinction coefficients using the procedure outlined in the instructions or use pierces online haba calculator. If the thermo scientific pierce biotin quantitation kit haba assay. 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. An adaptation of pierce s bca protein assay reagent kit product no.
The habaavidin premix is supplied in convenient noweigh microtube packaging, which eliminates the difficulties as sociated with weighing small quantities of reagent. Fluorescence biotin quantitation kit thermo fisher scientific. 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. For proteins of lower molecular weight or multiple biotin labels, less protein can be used.
After labeling, the extra sulfonhslcbiotin and sulfonhs byproduct were removed by gel filtration using zeba spin desalting columns. Bacterial endotoxin catalyzes theactivation of a proenzyme in the modified limulus amebocyte lysate lal. 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. Assessment of biotin interference with qualitative point. The p4 resin biogel p4 gel, medium was purchased from biorad laboratories hercules, ca. Biotin quantitation with quanttag kit from vector laboratories. Thermo scientific pierce biotin quantitation kit includes aliquots of haba reagent. Biotin 3 end dna labeling kit from pierce biotechnology. The thermo scientific fluorescence biotin quantitation kit requires only 10.
Pierce biotin quantitation kit thermo fisher scientific. Thermo scientific pierce fluorescence biotin quantitation kit. Biotin quantitation reagents and kits fisher scientific. 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. 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. Fluorescent dyes, biotins and streptavidins, sensolyte biotin quantitation kit fluorimetric. Recombinant antigen tpp15biotin for treponema pallidum. Surelinktm chromophoric biotin labeling kit for labeling and. Dec 08, 2006 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. Ingel detection of biotin protein conjugates with a green fluorescent streptavidin probe. Ample materials are provided to perform 25 assays in a cuvette format or 120 assays in a 96well format. Thermo scientific pierce biotin quantitation kit available on gsava contract for federal government customers only. 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.
Pierce bca protein assay kit brigham young university. Thermo scientific pierce fluorescence biotin quantitation kit requires only ten microliters of sample to accurately measure the biotinylation level of labeled. Haba 24hydroxyazobenzene benzoic acid and avidin are the basis of the kit. The resulting biotin conjugates were used in elisa, western blot. 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 fluorescence biotin quantitation kit is requires only 10 l of sample. Amplite colorimetric biotin quantitation kit aat bioquest. Surelinktm chromophoric biotin labeling kit for labeling and quantitation of. The activated proenzyme then catalyzes the splitting of p.
Many of the biotin labeling kits from pierce use an nhsbiotin reagent without the chromogenic fluorophore. It isnt always easy to find a trusted source of lab equipment and chemicals like this. The habaavidin premix is supplied in convenient thermo scientific noweigh microtube packaging, which. The molar ratio of biotin to her2 ecd was determined to be 9. The ez biotin quantitation kit and the streptavidin were purchased from pierce thermo fisher scientific, rockford, il.
The pierce bca protein assay is a detergentcompatible formulation based on bicinchoninic acid bca for the colorimetric detection and quantitation of total protein. 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. Determination of drugtoantibody ratio for antibodydrug. The assay utilizes haba 24hydroxyphenylazobenzoic acid, a reagent that shows dramatic spectral changes when bound to avidin. 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. Accuorange protein quantitation kit is a highly sensitive fluorescencebased assay for quantifying purified protein samples in 96well format. Two micrograms of biotinylated her2 ecd were immobilized on each assaymap saw cartridge using the agilent assaymap bravo controlled. If the pierce biotin quantitation kit will be performed to determine the. Compatibility chart for bca kit the amount listed is the maximum amount of material allowed in the protein sample without causing a noticeable interference, when 20 ul protein sample is used for bca assay as described in the standard assay. Assessment of biotin interference with qualitative pointof. Absorbance at 354 nm was more reproducible than hababased quantitation, could be performed with lower concentrations of sample, and was nondestructive.
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. Haba dye reagent and biotinylated protein standard for convenient colorimetric determination of biotinylation levels in antibodies and other proteins. The thermo scientific pierce biotin quantitation kit contains a premix of haba and avidin and a biotinylated horseradish peroxidase hrp positive control. The extent of biotin labeling depends on the distribution of amino groups on the protein, protein. Biotin quantitation kit colorimetric k811 biovision, inc. 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 biotinylation levels in the labeled her2 ecd were estimated using a pierce biotin quantitation kit, per the.
The kit is shipped on dry ice and storage at 20 c, protected from light, is recommended. Fluoreporter biotin quantitation assay kit for biotinylated proteins f30751 figure 1. Thermo scientific haba 4hydroxyazobenzene2carboxylic. After labeling, the extra sulfonhslc biotin and sulfonhs byproduct were removed by gel filtration using zeba spin desalting columns. Amount of biotin can be calculated directly from the decreased absorbance at 500nm complexing with the haba dye a convenient, accurate method for determining the degree of biotinylation. 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. The fluorescence biotin quantitation kit is requires only 10l of sample. 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.
By contrast, the traditional colorimetric method requires 100l of sample, and another leading suppliers fluorescent assay kit requires 50l of sample. Biovision biotin quantitation kit colorimetric, k811100. The thermo scientific pierce biotin quantitation kit contains a premix of. Biotinylation yields were evaluated using either the pierce biotin quantitation kit following the. Noweigh format ezlink sulfonhslcbiotin is shipped with an ice pack. The 99 mo99m tc generator was from perkin elmer life science inc boston, ma. The convenient assay can be performed either in a cuvette or in a microplate. This product is manufactured by expedeon, an abcam company, and was previously called lightninglink rapid biotin type b labeling kit. Determine biotinylation levels in labeled antibodies and other proteins using this fluorescent assay reagent and haba dye for rapid and sensitive measurement. Biotin vitamin h is an important biomolecule that has been widely used in biotinylation reactions to label antibodies or other proteins of interest. Determine the molar ratio of biotin incorporated into a. Sulfonhs biotin reagents see related thermo scientific products do not readily permeate cell membranes and are commonly used for specifically labeling the cell surface. By contrast, nhs biotin reagents are membrane permeable and may be used to biotinylate proteins inside intact cells.
Determine the molar ratio of biotin incorporated into a protein using the habaavidin method. Hello select your address best sellers customer service new releases find a gift whole foods registry gift cards sell customer service new. The sensitivity of biotin quantitation chromophoric biotin absorbance is 5 to 10fold greater than the sensitivity of the. The habaavidin premix is supplied in convenient thermo scientific noweigh microtube packaging, which eliminates the difficulties associated with. Biotin quantitation kit colorimetric k811 biovision. When haba is bound to streptavidin, addition of free biotin or biotinylated biomolecules results in its displacement in the reaction. 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. Ez biotin quantitation kit from pierce biocompare product. The pierce micro bca protein assay reagent kit is a detergentcompatible bicinchoninic acid formulation for the colorimetric detection and quantitation of total protein.
Protein analysis, modification and interaction biotinylation kits. 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. Biotin conjugation kit easy biotin labeling ab201796.
Many variations of this procedure exist in the literature. The tdt prefers to label singlestranded dna, but can also label doublestranded dna with blunt or 3 overhangs, but at a lower efficiency. Colorimetric biotin assay kit sufficient for 200 colorimetric. Surelinktm chromophoric biotin labeling kit for labeling. 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. Pn ez biotin quantitation kit, thermo scientific pierce the thermo scientific pierce high capacity streptavidin and neutravidin chromatography cartridges. What is the difference between the trilink biotin labeling. Incompatible substances amount compatible buffer systems nacetylglucosamine 10 mm in pbs, ph 7. If the drug requires sulfotag conjugation, label the drug following the protocol outlined in the msd. Abcam s biotin quantitation kit colorimetric ab185441 is based on the differential binding of streptavidin to biotin and a dye, 24hydroxyazobenzene benzoic acid haba. This kit can be used to determine accurately the labeling efficiency of biotinlabeled molecules. Ingel detection of biotinprotein conjugates with a green.
Furthermore, detection of protein biotinylation using stvgfp was easily detectable with 75 pmol poiavitag under nonoptimal uv excitation conditions, comparing favourably with the linear range of a commercial fluorescence biotin quantitation kit pierce, which has a. Our kit is more accurate than the other suppliers fluorescent biotin assay brand i. 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. Thermo scientific pierce biotin quantitation kit ez biotin. Pierce lal chromogenic endotoxin quantitation kit, 50 test 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 pierce haba is 4hydroxyazobenzene2carboxylic acid, a simple reagent that enables spectrophotometric colorimetric estimation of biotinylation levels of labeled proteins and other molecules.
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