Glutathione S-Transferase Fluorescent Kit

Details for Product No. ABIN577650, Supplier: Log in to see
Antigen
  • GST
  • GST 13-13
  • GST13
  • GST13-13
  • GSTK1-1
  • hGSTK1
  • GSTS
  • GSTS1-1
  • PGD2
  • PGDS
  • H-PGDS
  • Ptgds2
  • 1500002K10Rik
  • Gst
  • glutathione S-transferase kappa 1
  • hematopoietic prostaglandin D synthase
  • microsomal glutathione S-transferase 1
  • GSTK1
  • HPGDS
  • Hpgds
  • Mgst1
Minimum Detection Limit
< 100 µU
Application
Biochemical Assay (BCA)
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Purpose The DetectX® Glutathione S-Transferase Fluorescent Activity kit is designed to quantitatively measure the activity of GST present in a variety of samples.
Brand DetectX®
Sample Type Serum, Plasma, Urine, Cell Lysate
Detection Method Fluorometric
Specificity Sample Types validated: Serum, Plasma and Cell Lysates
Sensitivity 2.70 mU/mL
Characteristics The Glutathione S-Transferase (GST) Fluorescent Activity kit is designed to quantitatively measure the activity of GST. A GST standard is provided to generate a standard curve for the assay. 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. In the end point mode, mixing the sample or standard with the supplied Detection Reagent and GSH yields a fluorescent product which is read at 460nm in a fluorescent plate reader with excitation at 390nm. For rate determination, the intensity at 460nm is read periodically and plotted versus time. 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. 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 to 11), the neutral mu (pI 5 to 7) and acidic pi (pH <5) classes are populated by additional subclasses, each isozyme displaying differential specificity for given electrophilic molecules. Given its pivotal role in ameliorating oxidative stress/damage, GST activity has been repeatedly investigated as a biomarker for arthritis, asthma, COPD, and multiple forms of cancer, as well as an environmental marker. Examination of GST isoforms and activity in human cancers, tumors and tumor cell lines has revealed the predominance of the acidic pi class. Furthermore, this activity is thought to substantially contribute to the innate or acquired resistance of specific neoplasms to anticancer therapy.
Components Black half Area 96 well Plate
Material not included Repeater pipet with disposable tips capable of dispensing 25 μL.
Fluorescence 96 well plate reader capable of reading fluorescent emission at 460 nm, with excitation at 390 nm.
Set plate parameters for a 96-well Corning Costar 3686 plate. Software for converting raw relative fluorescent unit (FLU) readings from the plate reader and carrying out four parameter logistic curve (4PLC) fitting.
Alternative Name Glutathione S-Transferase (GST ELISA Kit Abstract)
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, 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 <5) classes are populated by additional subclasses, each isozyme displaying differential specificity for given electrophilic molecules2. Given its pivotal role in ameliorating oxidative stress/damage, GST activity has been repeatedly investigated as a biomarker for arthritis, asthma, COPD, and multiple forms of cancer, as well as an environmental marker3-7. Examination of GST isoforms and activity in human cancers, tumors and tumor cell lines has revealed the predominance of the acidic pi class. Furthermore, this activity is thought to substantially contribute to the innate or acquired resistance of specific neoplasms to anticancer therapy8,9
Application Notes 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.
Samples that are not clear or that contain visible particulate should be centrifuged prior to using.
GST activity varies across tissues and species, however we expect this kit to measure GST activity from sources other than human.
The end user should evaluate recoveries of GST activity in samples from other species being tested.
Assay Time 1 h
Plate 96 wells
Protocol 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.
Sample Preparation

Any samples requiring larger dilutions or with GST activities outside the standard curve range should be diluted further with Assay Buffer to obtain readings within the standard curve. urine Samples Samples that are not clear or that contain visible particulate should be centrifuged prior to using. Urine samples should be diluted ≥ 1:2 in Assay Buffer by adding one part of urine to one part of Assay Buffer. Sample values should be normalized for urinary volume to urinary creatinine levels. Our Urinary Creatinine Detection Kits, K002-H1 and K002-H5, allow simple, safe and accurate determination of urinary creatinine levels. Serum and

Restrictions For Research Use only
Precaution of Use As with all such products, this kit should only be used by qualified personnel who have had laboratory safety instruction.
The complete insert should be read and understood before attempting to use the product.
Dimethyl sulfoxide is a powerful aprotic organic solvent that has been shown to enhance the rate of skin absorption of skin-permeable substances.
Wear protective gloves when using the solvent especially when it contains dissolved chemicals.
Handling Advice ACtivity StAndArdizAtion The Glutathione S-Transferase standard used in this kit has been calibrated using an enzymatic method adapted from reference 1.
Storage 4 °C
Storage Comment All components of this kit should be stored at 4°C until the expiration date of the kit. DMSO, when stored at 4°C, will freeze. Can be stored tightly capped at room temperature.
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