Invitrogen™

Glutathione S-Transferase Fluorescent Activity Kit

Catalog number:  EIAGSTF

 Related applications:

Protein Assays and Analysis

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Description

The Glutathione S-Transferase Activity research-use-only kit is a fluorescent activity assay designed for the quantification and detection of glutathione S-transferase activity in serum, plasma, urine and cell lysates.

This complete, ready-to-use kit includes black 96-well plate(s), glutathione S-transferase standard (10 U/mL), glutathione S-transferase substrate, and other components to perform the assay. A 96-well microplate reader capable of reading the fluorescent emission at 460 nm, with excitation at 370-410 nm, is required for use of this kit.

Performance characteristics
• Assay type: fluorescent activity kit
• Sample types: serum, plasma, urine, and cell lysates
• Sensitivity: 2.70 mU/mL
• Standard curve range: 7.8 mU/mL–500 mU/mL
• Reactivity: species independent

Background
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 glutathione1. Human GSTs are encoded by 5 gene families, expressed 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<5) classes are populated by additional subclasses, each isozyme displaying differential specificity for given electrophilic molecules. This assay has been validated for human urine, serum, EDTA, heparin plasma, toadfish liver (Opsanus tau), and oyster hemolymph samples. Most cell lysates should also be compatible. GST activity varies across tissues and species, so this kit should measure GST activity from sources other than human.

Assay principle
The Glutathione S-Transferase Fluorescent 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 that 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.

Related links
Learn more about ELISA kits
Learn more about other immunoassays

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

Specifications

Immunoassay Kit Format: Activity Kit
Target (Kit Named): Glutathione S-Transferase
For Use With (Equipment): Microplate Reader
Product Size: 96 tests
Shipping Condition: Approved for shipment on Wet or Dry Ice

Contents & storage

96-well plate(s), standard, detection reagent, assay buffer, substrate, and lot-specific technical data sheet.

Store at –20°C.

Documents

Manuals & protocols