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MAK151

Sigma-Aldrich

Fluorometric Thiol Assay Kit

sufficient for 200 fluorometric tests

Synonym(s):

Thiol Fluorescent Assay Kit

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About This Item

EC Number:
200-664-3
UNSPSC Code:
12161503
NACRES:
NA.84
Pricing and availability is not currently available.

usage

sufficient for 200 fluorometric tests

detection method

fluorometric

relevant disease(s)

endocrinological disorders, diabetes; cancer

storage temp.

−20°C

General description

Thiol groups, found as free cysteine, glutathione (GSH), and cysteine residues in proteins, are involved in many biological processes. The disulfide bonds generated when the thiol groups of two cysteine residues are oxidized contribute to the tertiary or quatenary structure of a protein.

Application

Fluorometric Thiol Assay Kit has been used to determine the lipid peroxidation expressed as thiols in samples.[1]

Suitability

This kit is suitable for the detection of thiols in a variety of different biological samples including plasma, urine, and cell extracts. This kit is not optimized for measuring thiol groups in proteins.

Principle

The Fluorometric Thiol Quantitation Kit provides an ultrasensitive assay to quantitate the thiol content of small molecules. The proprietary thiol sensor generates a strongly fluorescent adduct (λex = 490/λem = 520 nm) upon reacting with a thiol-containing compound. In addition, both absorption and emission spectra of the thiol adduct are pH-independent, making this assay kit highly robust. The kit can detect as little as 1 picomole of cysteine or GSH in a 100 μL assay volume (10 nM). The assay can be performed conveniently in either a 96 or 384 multiwell plate format.

Storage Class Code

10 - Combustible liquids

WGK

WGK 3

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Certificates of Analysis (COA)

Lot/Batch Number
3510891
304162
3320703
2781663
2391623

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D Scott Smith et al.
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Xiao-Dong Gong et al.
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Protein sumoylation is one of the most important post-translational modifications regulating many biological processes (Flotho A & Melchior F. 2013. Ann Rev. Biochem. 82:357-85). Our previous studies have shown that sumoylation plays a fundamental role in regulating lens differentiation (Yan
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Questions

1–4 of 4 Questions  

  1. 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

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  2. 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?

  3. Can this kit be used to quantify cysteines from proteins? If so, do I need to reduce disulfide bonds first? Also, does maleimide interfere with the assay?

    1 answer

    1. This kit detects reduced thiols such as reduced Glutathione (GSH) and Cysteine. In addition, the product may also be used to detect SH groups in the protein, provided that a standard with known SH groups is used, such as BSA (0.5-1 GH/molecule). The possible interference of Meleimide has not been investigated.

      Helpful?

  4. Would MAK151 detect reduced thiols like GSH, Cysteine, and NAC, or does it detect total thiol? Additionally, does it detect total protein-SH or protein-S-S protein?

    1 answer

    1. MAK151 is capable of detecting reduced thiols like GSH, Cysteine, etc. It can also be used to detect SH groups in proteins, provided that known SH groups of the protein standard, such as BSA (0.5-1 GH/molecule), are available.

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