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CGP1

Sigma-Aldrich

Glutathione Peroxidase Cellular Activity Assay Kit

Sufficient for 100 colorimetric tests

Synonym(s):

Cellular glutathione peroxidase, c-GPx

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About This Item

UNSPSC Code:
12161503
NACRES:
NA.32
Pricing and availability is not currently available.

Quality Level

200

usage

sufficient for 100 tests

shelf life

Kit is stable for 24 months when unopened.

detection method

colorimetric

shipped in

wet ice

storage temp.

−20°C

General description

The Glutathione Peroxidase Cellular Activity Assay kit is used to measure glutathione peroxidase in tissue extracts. The assay is based on the oxidation of glutathione (GSH) to oxidized glutathione (GSSG). This is catalyzed by GPx coupled to the recycling of GSSG back to GSH utilizing glutathione reductase and NADPH. The decrease in NADPH absorbance measured at 340 nm during the oxidation of NADPH to NADP is indicative of glutathione peroxidase activity since the enzyme is the rate-limiting factor of the coupled reactions.

Suitability

Suitable for the measureent of glutathione peroxidase in tissue extracts

Principle

The Glutathione Peroxidase Cellular Activity Assay kit is used to measure glutathione peroxidase in tissue extracts. The assay is based on the oxidation of glutathione (GSH) to oxidized glutathione (GSSG). This is catalyzed by GPx coupled to the recycling of GSSG back to GSH utilizing glutathione reductase and NADPH. The decrease in NADPH absorbance measured at 340 nm during the oxidation of NADPH to NADP is indicative of glutathione peroxidase activity since the enzyme is the rate-limiting factor of the coupled reactions.

Other Notes

This kit was tested with rabbit reticulocyte lysate and with glutathione peroxidase enzyme.

Kit Components Only

Product No.
Description
  • tert-Butyl hydroperoxide 1 mL

related product

Product No.
Description
Pricing

Signal Word

Danger

Hazard Classifications

Acute Tox. 2 Inhalation - Acute Tox. 3 Dermal - Acute Tox. 4 Oral - Aquatic Chronic 2 - Eye Dam. 1 - Flam. Liq. 3 - Muta. 2 - Org. Perox. F - Skin Corr. 1C - Skin Sens. 1 - STOT SE 3

Target Organs

Respiratory system

Storage Class Code

5.2 - Organic peroxides and self-reacting hazardous materials

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Certificates of Analysis (COA)

Lot/Batch Number
0000397879
0000397879
0000182208
078M4136V
11180343

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V R Arruda et al.
Neoplasma, 43(2), 99-102 (1996-01-01)
Red cell antioxidant enzymes have been recently studied in malignant lymphomas and the results are controversial. Hairy cell leukemia is a rare chronic lymphoproliferative disorder originating probably in a pluripotent stem cell. In the present study, glutathione peroxidase (Gpx), reduced
Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase.
D E Paglia et al.
The Journal of laboratory and clinical medicine, 70(1), 158-169 (1967-07-01)
B L Gupta et al.
Biochemistry and molecular biology international, 46(6), 1145-1152 (1999-01-19)
Experimentally induced diabetic rats were treated separately with insulin and vanadate. The activities of hexokinase (HK) and glucose-6-phosphate dehydrogenase (G-6PDH) were increased in reticulocyte hemolysate isolated from the diabetic rats and were restored to normal levels by insulin. The restoration
International Committee for Standardization in Haematology: recommended methods for red-cell enzyme analysis.
E Beutler et al.
British journal of haematology, 35(2), 331-340 (1977-02-01)
Ian M Scott et al.
Pesticide biochemistry and physiology, 141, 9-17 (2017-09-16)
Plant elicitors can be biological or chemical-derived stimulators of jasmonic acid (JA) or salicylic acid (SA) pathways shown to prime the defenses in many crops. Examples of chemical elicitors of the JA and SA pathways include methyl-jasmonate and 1,2,3-benzothiadiazole-7-carbothioate (BTH
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Questions

1–10 of 10 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. "Does A340/MIN refer to the absorbance value per minute, or is it the rate of decrease per minute?"

    1 answer

    1. The CGP1 assay measures the decrease in NADPH absorbance at 340 nm during the oxidation of NADPH to NADP+. This is a kinetic assay and the A340/MIN means absorbance per minute. Please see step 4 on page 4 of the technical bulletin for the kinetic program instructions. The number of readings being 6 for an interval time of 10 seconds which means data is for a minute. NADP+ does not have any absorbance at 340 nm and NADPH does. Thus, the absorbance will decrease in a sample with glutathione peroxide as all substrates, cofactors, enzymes are present for this conversion to occur.

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  4. Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be used with Serum samples?

    1 answer

    1. The Glutathione Peroxidase Cellular Activity Assay Kit (Product number: CGP1) is designed to measure Glutathione Peroxidase activity in cell lysates and tissue homogenates. This kit has not been validated for use with serum samples.

      Please see product MAK437, Glutathione Peroxidase Assay Kit, which can be used for serum samples:
      https://www.sigmaaldrich.com/product/sigma/mak437

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  5. Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be run in a 96 well plate?

    1 answer

    1. Yes, but you would need to have a plate reader that has a kinetic function as this is a kinetic assay.

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  6. What is the Department of Transportation shipping information for this product?

    1 answer

    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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  7. How should I prepare my sample for analysis with Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?

    1 answer

    1. The cells must be lysed before assaying with this kit.  The best way to perform lysis is to incubate the cells with hypotonic buffer (at least 2 volumes and better with 3-4 volumes, depending on the expected level of peroxidase in these cells). Let them swell and homogenize with a Dounce homogenizer. Centrifuge the homogenate at 10,000 × g and use the supernatant for the assay.  If a detergent is used for lysis, only use a peroxide-free detergent and follow the instructions in the Bulletin: "Nonionic detergents such as TWEEN and TRITON X-100 that contain high levels of endogenous peroxides will raise the apparent activity. If these detergents are vital to the extraction of the proteins of interest, a low peroxide detergent should be used such as Product Codes X-100-PC (TRITON X-100), P 6585 (TWEEN 20), or P 8192 (TWEEN 80)".

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  8. What dilution should I use for my sample when using Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?

    1 answer

    1. In general, when human erythrocytes were assayed for the enzyme, the sample was diluted 10-fold (15 mg peroxidase per mL) and a 20 μl sample gave a decrease of 0.032 OD340 per minute. Too much enzyme will underestimate the apparent activity of the sample. When rabbit reticulocytes were assayed at a 10-fold dilution (10 mg peroxidase per mL), 20 μl gave a decrease of 0.065 OD340 per minute.

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  9. What type of readings do I get with Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?

    1 answer

    1. There is a decrease in absorbance during the assay. The OD values decrease with the increase in reaction time. The slope of the reduction in OD (delta OD per minute) is the value used to determine the enzyme units.

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  10. Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be used with tissue samples?

    1 answer

    1. We have not tested the CGP1 assay with tissue samples. However, we suggest that the customer perform a small-scale pilot experiment using our CelLytic-MT lysis buffer (Product No. C3228). The buffer enables efficient extraction of tissue proteins. It does not contain any of the interfering agents that are listed on the kit bulletin.

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