Thermo Scientific™ Pierce™ Fluorescence Biotin Quantitation Kit
Determine biotinylation levels in labeled antibodies and other proteins using this fluorescent assay reagent and HABA dye for rapid and sensitive measurement.
Manufacturer: Thermo Scientific™ 46610
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.
- 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
Assay buffer, HABA-avidin fluorescent reporter solution, and biocytin control
Black opaque 96-well microplate and a fluorescence plate reader (excitation 494nm; emission 520nm)
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.
|Antibodies (General), Proteins (General)|
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