Pierce™ Biotin Quantitation Kit - FAQs

View additional product information for Pierce™ Biotin Quantitation Kit - FAQs (28005)

3 product FAQs found

How can I determine the amount of biotinylation on my ligand?

An assay with HABA dye (Cat. No. 28010 or 28005) can be used to determine the molarity of biotin in solution.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can you provide the shelf life for Pierce Biotin Quantitation Kit (28005)?

Pierce Biotin Quantitation Kit is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended (4 degrees C). Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf).

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

How can I determine the degree of protein labeling?

To quantitate biotin, we offer two kits as follows:

Biotin Quantitation Kit (Cat. No. 28005): With this kit, a solution containing the biotinylated protein is added to a mixture of HABA reagent (4’-hydroxyazobenzene-2-carboxylic acid and avidin. Because of its higher affinity for avidin, biotin displaces the HABA and the absorbance at 500nm decreases proportionally.

Fluorescence Biotin Quantitation kit (Cat. No. 46610): 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 must less sample volume than the microplate colorimetric HABA assay and is much more sensitive

To determine the dye-to-protein ratio after fluorophore conjugation, absorbance readings of the protein:dye conjugate are taken and the molar ratio can then be calculated. Please go to this Tech Tip (https://tools.thermofisher.com/content/sfs/brochures/TR0031-Calc-FP-ratios.pdf")for more information

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.