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Thermo Scientific™ Pierce™ Biotin Quantitation Kit

Catalog No. PI28005
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12 Tests
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PI28005 12 Tests
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Catalog No. PI28005 Supplier Thermo Scientific™ Supplier No. 28005
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Description

HABA dye reagent and biotinylated protein standard for convenient colorimetric determination of biotinylation levels in antibodies and other proteins.

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.

Determine the molar ratio of biotin incorporated into a protein using the HABA-avidin method. The HABA dye (4'-hydroxyazobenzene-2-carboxylic acid) binds to avidin to produce a yellow-orange colored complex that absorbs at 500nm. Free biotin present in solution with this avidin-HABA complex will displace the HABA dye and cause the absorbance to decrease.The amount of biotin present can be calculated directly from the decreased absorbance at 500nm. The Thermo Scientific EZ Biotin Quantitation Kit contains pre-measured doses of the Avidin-HABA mixture and biotinylated horseradish peroxidase positive control to simplify reagent preparation and minimize the amount of waste generated. The convenient assay can be performed either in a cuvette or in a microplate.

Highlights:

  • HABA-avidin complex can be used over a wide range of pH and salt concentrations
  • Amount of biotin can be calculated directly from the decreased absorbance at 500nm complexing with the HABA dye

Eliminate the need for cumbersome assay equations with a convenient HABA calculator found at www.piercenet.com/HABA/habacalc.cfm

Includes:

HABA/Avidin Premix, 24 tubes; Biotinylated HRP, 12 tubes

Specifications

Product Type Biotin Quantitation Kit
Quantity 12 Tests
Content And Storage Sufficient For: Testing biotinylation of 12 protein samples
• HABA-Avidin Premix, 24 tubes
• Biotinylated Horseradish Peroxidase, 5 mg

Store at 4°C.
Product Line Pierce
Label or Dye Biotin
Labeling Target Antibodies, Proteins

Frequently Asked Questions (FAQs)

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.
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).
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

    For Research Use Only. Not for use in diagnostic procedures.