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Key Documents

MAK171

Sigma-Aldrich

Colorimetric Biotin Assay Kit

sufficient for 200 colorimetric assays

Synonym(s):

Biotin Quantitation Kit

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About This Item

EC Number:
200-399-3
UNSPSC Code:
12352203
NACRES:
NA.84
Pricing and availability is not currently available.

detection method

colorimetric

relevant disease(s)

dermatological diseases; neurological disorders

shipped in

dry ice

storage temp.

−20°C

Related Categories

General description

Biotinylation is the process of labelling a molecule with a biotin molecule to take advantage of the strong binding and high specificity of biotin and streptavidin for purification or detection applications. Biotinylation is typically used to label proteins or nucleic acid based targets for these applications.[1] Quantitation of biotinylation can be used to optimize the labelling of a given target. Levels of biotinylation can effect detection sensitivity or functionality of a purification product.[2]

Suitability

Suitable for estimating the molar ratio of biotin to protein in biotin-protein conjugates or for quantitating biotin concentration in a solution

Principle

This Colorimetric Biotin Quantitation Kit provides a convenient method for estimating the molar ratio of biotin to protein in biotin-protein conjugates or for quantitating biotin concentration in a solution. The assay utilizes HABA (4′-hydroxyazobenzene-2-carboxylic acid), a reagent that shows dramatic spectral changes when bound to avidin. Biotin easily displaces HABA from the HABA/Avidin complex, resulting in a decrease of absorption at 500 nm.

Preparation Note

Storage and Stability: The kit is shipped on dry ice and storage at –20 °C, protected from light, is recommended.

Precautions and Disclaimer: This product is for R&D;use only, not for drug, household, or other uses. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices.

Storage Class Code

10 - Combustible liquids

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Certificates of Analysis (COA)

Lot/Batch Number
3511761
3321392
3180703
3511761
3321392

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High-throughput biotinylation of proteins.
High Throughput Protein Expression and Purification, 185-198 (2009)
Site-specific biotinylation of purified proteins using BirA.
Fairhead M and Mark H
Site-Specific Protein Labeling, 171-184 (2015)
Imre Gaspar et al.
Bio-protocol, 8(5), e2750-e2750 (2018-03-05)
Arrays of short, singly-labeled ssDNA oligonucleotides enable in situ hybridization with single molecule sensitivity and efficient transcript specific RNA capture. Here, we describe a simple, enzymatic protocol that can be carried out using basic laboratory equipment to convert arrays of

Questions

1–6 of 6 Questions  

  1. How can I determine the shelf life / expiration / retest date of this product?

    1 answer

    1. If this product has an expiration or retest date, it will be shown on the Certificate of Analysis (COA, CofA). If there is no retest or expiration date listed on the product's COA, we do not have suitable stability data to determine a shelf life. For these products, the only date on the COA will be the release date; a retest, expiration, or use-by-date will not be displayed.
      For all products, we recommend handling per defined conditions as printed in our product literature and website product descriptions. We recommend that products should be routinely inspected by customers to ensure they perform as expected.
      For products without retest or expiration dates, our standard warranty of 1 year from the date of shipment is applicable.
      For more information, please refer to the Product Dating Information document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/449/386/product-dating-information-mk.pdf

      Helpful?

  2. How is shipping temperature determined? And how is it related to the product storage temperature?

    1 answer

    1. Products may be shipped at a different temperature than the recommended long-term storage temperature. If the product quality is sensitive to short-term exposure to conditions other than the recommended long-term storage, it will be shipped on wet or dry-ice. If the product quality is NOT affected by short-term exposure to conditions other than the recommended long-term storage, it will be shipped at ambient temperature. As shipping routes are configured for minimum transit times, shipping at ambient temperature helps control shipping costs for our customers. For more information, please refer to the Storage and Transport Conditions document: https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/marketing/global/documents/316/622/storage-transport-conditions-mk.pdf

      Helpful?

  3. What are the instructions for running the kit with catalog number MAK171? Additionally, needs more information to understand the purpose of this kit and how to perform calculations if purchased.

    1 answer

    1. The Colorimetric Biotin Quantitation Kit with catalog number MAK171 offers a convenient method for estimating the molar ratio of biotin to protein in biotin-protein conjugates or for quantifying biotin concentration in a solution. The assay utilizes HABA [2-(4-Hydroxyphenylazo)benzoic acid], a reagent that exhibits significant spectral changes when bound to avidin. Biotin readily displaces HABA from the HABA/Avidin complex, leading to a decrease in absorption at 500 nm. The kit provides an optimal ratio of Avidin and HABA and is most effective for determining biotin concentration in the range from 2–16 mM. The assay can be carried out in a cuvette or microplate format.

      Helpful?

  4. Is it possible to use RIPA buffer as the lysis reagent with the Colorimetric Biotin Assay Kit, product number MAK171-1KT, to extract proteins and biotin from samples such as brain parenchymal tissues?

    1 answer

    1. RIPA buffer as the lysis reagent is acceptable, provided that the concentration of the RIPA buffer is consistent with that of the control.

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  5. Pourquoi la concentration du control positif n'est pas donnée ? Question de savoir si le test a bien fonctionné et la concentration de notre échantillon est juste.

    1 answer

    1. The positive control is a vial of 100uL of 100uM d-biotin.

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  6. Can I use it to measure serum samples?

    1 answer

    1. This product is suitable for biotin-containing samples. The range of detection for this kit is 2 - 16 uM. Biotin concentrations would need to be within this range. Please see the link below to review the product datasheet for this kit:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/116/046/mak171bul.pdf

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