Accurately measure biotinylation levels with a new highly sensitive fluorescent assay
The Thermo Scientific Fluorescence Biotin Quantitation Kit requires only 10 µl of sample to accurately measure the biotinylation level of biomolecules. This microplate-based assay is easy to perform by adding the Thermo Scientific DyLight 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 half the sample volume and is much more sensitive than the microplate colorimetric HABA assay.
The highly specific interaction of avidin with biotin is useful for designing nonradioactive purification and detection systems. Quantifying the degree of biotinylation is necessary for determining if biotinylation was successful and the amount of biotinylated molecule to use for a specific application. The Fluorescence Biotin Quantitation Kit contains a premix of fluorescent avidin with HABA and a biocytin standard. This fluorescent method is highly sensitive and accurate and requires only 10 µl of sample, whereas the traditional colorimetric method requires 100 µl of sample and Supplier I's fluorescent assay requires 50 µl of sample. Additionally, our kit more accurately quantitates biotin than Supplier I's fluorescent assay.
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; however, our new fluorescent method is more sensitive and accurate. The premix 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. Thermo Scientific Fluorescence Biotin Quantitation kit ordering information
1 Items Total 2,415.00 USD 
Accurately measure biotinylation levels with a new highly sensitive fluorescent assay