Thermo Scientific™ Pierce™ Fluorescence Biotin Quantitation Kit

Description

• Thermo Scientific™ Pierce Fluorescence Biotin Quantitation Kit requires only ten microliters of sample to accurately measure the biotinylation level of labeled antibodies and other biomolecules
• This microplate-based biotin assay is easy to perform by adding the supplied fluorescent reporter to the biotinylated samples and diluted biocytin standards
• The avidin fluoresces when the weakly interacting HABA (4'-hydroxyazobenzene-2-carboxylic acid) is displaced by the biotin
• The amount of biotin is determined by comparing the sample's fluorescence to the biocytin standard curve
• This assay requires much less sample volume than the microplate colorimetric HABA assay and is much more sensitive
• Highlights: Fast — requires only 5 minute incubation Economical — requires only 10μL of sample (approx
• 750ng of biotinylated IgG) Sensitive — linear working range of 10 to 60 picomoles of biotin Includes: Assay buffer, HABA-avidin fluorescent reporter solution, and biocytin control Requires: Black opaque 96-well microplate and a fluorescence plate reader (excitation 494nm; emission 520nm) Product Details: The highly specific interaction of avidin or streptavidin with biotin is the basis for many purification and detection systems
• Quantifying the degree of biotinylation is necessary for assessing if a biotin-labeling procedure was successful and what amount of biotinylated molecule to use for a specific application
• HABA (4'-hydroxyazobenzene-2-carboxylic acid) is a dye that weakly interacts with avidin and is commonly used in a colorimetric assay to quickly estimate the biotin-to-protein ratio
• The Fluorescence Biotin Quantitation Kit also uses HABA dye but is more sensitive and accurate
• A premix of fluorescent avidin with HABA (DyLight Reporter) is added to the solution containing the biotinylated sample
• Because of its higher affinity for avidin, biotin displaces the HABA, allowing the avidin to fluoresce
• The amount of biotin is measured in a microplate by comparing the fluorescence to a biocytin standard curve
• The Fluorescence Biotin Quantitation Kit is requires only 10μL of sample
• By contrast, the traditional colorimetric method requires 100μL of sample, and another leading supplier's fluorescent assay kit requires 50μL of sample
• Our kit is more accurate than the other supplier's fluorescent biotin assay.