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Stock availability
In Stock
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Kit s description
Quantitative fluorometric measurement of the activity of GST
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Protein target
Glutathione S-Transferase
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Scientific context
The Glutathione S-Transferase (GST) family of isozymes function to detoxify and neutralize a wide variety of electrophilic molecules by mediating their conjugation with reduced glutathione (1). Human GSTs are encoded by 5 gene families, expressing in almost all tissues as four cytosolic and one microsomal forms. Dividing the family by isoelectric points, the basic alpha (pI 8–11), the neutral mu (pI 5–7) and acidic pi (pH
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Primary research area
Cancer, Oxidative Stress
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Category
Assay Activity Kits
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Other name
γ-L-Glutamyl-L-cysteinylglycine Activity Kit (2S)-2-Amino-5-[[(2R)-1-(carboxymethylamino)-1-oxo- 3-sulfanylpropan-2-yl]amino]-5-oxopentanoic acid Activity Kit
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Brand name
StressXpress®
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Detection system
Fluorometric Assay
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Assay format
Direct Enzyme Activity Assay
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Main intended usage
Fluorometric assay used to quantitatively measure the activity of glutathione S-transferase in samples.
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Brief protocol
The Glutathione S-Transferase Activity kit is designed to quantitatively measure the activity of GST present in a variety of samples. A GST standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. The kit utilizes a non-fluorescent molecule that is a substrate for the GST enzyme which covalently attaches to glutathione (GSH) to yield a highly fluorescent product. Mixing the sample or standard with the supplied Detection Reagent and GSH and incubating at room temperature for 30 minutes yields a fluorescent product which is read at 460 nm in a fluorescent plate reader with excitation at 390 nm. The activity of the GST in the sample is calculated, after making a suitable correction for any dilution of the sample, using software available with most fluorescence plate readers. We have provided a 96 well plate for measurement but this assay is adaptable for higher density plate formats. The end user should ensure that their HTS black plate is suitable for use with these reagents prior to running samples.
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Kit contents
Black Half Area 96 Well Plate, Glutathione S-Transferase Standard, GST Detection Reagent, Dry DMSO, Assay Buffer, Glutathione (GSH)
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Assay precision
Intra Assay Precision:Four serum samples were diluted 1:2 with Assay Buffer and run in replicates of 16 in an assay. The mean and precision of the calculated GST activities were: Sample 1- 315.9 mU/mL, 4.6% CV Sample 2- 221.2 mU/mL, 5.6% CV Sample 3- 88.2 mU/mL, 4.2% CV Sample 4- 22.7 mU/mL, 6.6% CV Inter Assay Precision: Four serum samples were diluted 1:2 with Assay Buffer and run in duplicates in twenty assays over multiple days by four operators. The mean and precision of the calculated GST activities were: Sample 1- 291.7 mU/mL, 12.6% CV Sample 2- 218.5 mU/mL, 11.0% CV Sample 3- 89.6 mU/mL, 10.4% CV Sample 4- 23.0 mU/mL, 15.9% CV
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Assay cross reactivity
Human, Mollusk, Fish
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Samples to be used with this kit
Urine, Serum, EDTA Plasma, Heparin Plasma, Cell Lysates
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Maximum samples to be used with this kit
40 samples in duplicate
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Assay duration
30 Minutes
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Kit s sensitivity
2,70 mU/ml
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Assay detection limit
7,81 - 500 mU/ml
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Storage recommendations
4°C
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Shipping recommendations
Blue Ice
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Bibliography
1. Habig, W, et al. J. Biol.Chem. 1974 249(22):7130-7139. 2. Cook, JA, et al. Cancer Res. 1991 51:1606-1612. 3. Dalle-Donne, I, et al. 2006 52(4):601-623. 4. Surapneni, KM & VSC Gopan. Ind.J.Clin.Biochem. 2008 23(1):41.44. 5. Mohan, SK & O Venkataramana. Ind.J.Med. Sci. 2007 61:9-14. 6. Ferrandina, G., et al. Ann. Oncol. 1997 8:343-350. 7. Otitoju, O & Onwarah, INE. African J. Biotech. 2007 6(12)1455-1459. 8. Shea, TC, et al. Cancer Res. 1988 48:527-533. 9. Shea, TC, et al. Cancer Res. 1990 50:6848-6853.
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Release date
17-Sep-2014
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PubMed number
Refer to PubMed
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Tested applications
No data available / currently testing
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Tested reactivity
No data available / currently testing
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Resources available upon request
Kit Booklet, MSDS
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Product image link
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Product image legend
Continuous Kinetic Assay Typical Standard Curve for Glutathione S-Transferase Activity Kit (Enzyme Activity Assay) StressXpress® - SKT-203. Assay Type: Direct. Detection Method: Fluorometric Assay. Assay Range: 7.81 - 500 mU/ml. | End-Point Assay Typical Standard Curve for Glutathione S-Transferase Activity Kit (Enzyme Activity Assay) StressXpress® - SKT-203. Assay Type: Direct. Detection Method: Fluorometric Assay. Assay Range: 7.81 - 500 mU/ml. | Linearity was determined by taking Jurkat cell lysates at 10 x 106 and 0.8 x 106 cells/mL and mixing in the ratios given below. The measured activities were compared to the expected values based on the ratios used. Graph of the Continuous Kinetic Assay Typical Standard Curve for the Glutathione S-Transferase Activity Kit StressXpress - SKT-203 | Graph of the End-Point Assay Typical Standard Curve for the Glutathione S-Transferase Activity Kit StressXpress - SKT-203 | Graph of the Linearity Recovery for the Glutathione S-Transferase Activity Kit StressXpress - SKT-203
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Warnings
Non-hazardous
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Country of production
Canada
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Total weight kg
2
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Net weight g
300
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