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Oxidative dammage

 

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 Assays for Detection of Oxidative Damage

Superoxide Dismutase (SOD) - Catalyzes the dismutation of the superoxide
radical (O2
-) into hydrogen peroxide (H202) and elemental oxygen (O2), and as
such, provides an important defense against the toxicity of superoxide radical.
Superoxide ions, generated from the conversion of xanthine to uric acid and
hydrogen peroxide by xanthine oxidase, converts NBT to NBT-diformazan. NBTdiformazan
absorbs light at 550 nm. SOD reduces the superoxide ion
concentration and thereby lowers the rate of NBT-diformazan formation. The kit
contains the proper lysis buffer and the reagents needed for 100 experimental
tests, 50 positive controls, and 50 negative controls.
Glutathione Reductase is an enzyme which catalyzes the reduction of oxidized
glutathione (GSSG) to reduced glutathione (GSH). GSH is an important
intracellular antioxidant that reacts with oxygen free radicals and organic
peroxides and acts as a substrate for other detoxification enzymes such as
glutathione peroxidase and glutathione S-transferase. The activity of glutathione
reductase is an important measure of the antioxidant status of the cell.
Glutathione reductase requires NADPH for its activity, resulting in the reduction
of GSSG to GSH and the corresponding oxidation of NADPH to NADP. Trevigen’s
Glutathione Reductase Assay takes less than 6 minutes to perform and is based
on the measurement of the loss in absorbance at 340 nm due to the conversion
of NADPH to NADP+.
Glutathione Assay Kit utilizes an optimized enzymatic recycling method, using
Glutathione reductase, for the quantification of Glutathione. The sulfhydryl group
of Glutathione reacts with DTNB (5,5’-dithiobis-2-nitrobenzoic acid, Ellman’s
reagent) and produces a yellow colored 5-thio-2-nitrobenzoic acid (TNB). The
mixed disulfide, GSTNB (between GSH and TNB) that is concomitantly produced,
is reduced by Glutathione reductase to recycle the GSH and produce more TNB.
The rate of TNB production is directly proportional to this recycling reaction
which is in turn directly proportional to the concentration of GSH in the sample.
Measurement of the absorbance of TNB at 405 or 414 nm provides an accurate
estimation of Glutathione in a sample.
Glutathione Peroxidase is a tetramer of four identical subunits, with a
molecular weight of 84,000. It requires selenium as a cofactor and contains a
selenocysteine amino acid residue in the active site of each monomer that
participates in the actual mechanism of the enzyme. Glutathione peroxidase
(GP) is found in mammalian cells and helps to prevent lipid peroxidation of cell
membranes by consuming free peroxide in the cell. Trevigen’s Glutathione
Peroxidase Assay Kit can be used to measure glutathione-dependent
peroxidases in plasma, erythrocyte lysates, tissue homogenates, and cell
lysates.
 

Features Benefits
96-well microplate formats for Glutathione
Reductase and Superoxide Dismutase Kits
High-throughput assay formats
Unlike other assay kits for SOD, Trevigen’s SOD
Kit is not greatly disturbed by trace metals
Improved experimental results
Glutathione Kit includes procedure for
determining total, oxidized and reduced
glutathione
Ability to test for various forms of
glutathione
Fast and efficient assays Each SOD assay sample takes only 5
minutes; Each glutathione reductase
sample takes only 6 minutes
Easy-to-use kit format All necessary reagents for fast,
consistent results
Ordering Information:
Oxidative Damage Assay Kits
7500-100-K SOD Assay Kit, 100 reactions
7512-100-K Glutathione Peroxidase Assay Kit, 100 tests
7511-100-K Glutathione Assay Kit, 100 tests
7510-100-K Glutathione Reductase Assay, 100 reactions


Oxidative Damage Enzymes
4045-05K-EB E. coli Endonuclease III Enzyme & Buffer, 5000 Units
4045-01K-EB E. coli Endonuclease III Enzyme & Buffer, 1000 Units
4060-05K-EB E. coli Endonuclease VIII Enzyme & Buffer, 5000 Units
4060-01K-EB E. coli Endonuclease VIII Enzyme & Buffer, 1000 Units
4040-500-EB E. coli Fpg Enzyme & Buffer, 2500 Units
4040-100-EB E. coli Fpg Enzyme & Buffer, 500 Units


Now Available: 8-oxo-dG Elisa Kit for detection and quantification of 8-
OHdG in urine and serum samples
Cat# 4370-096-K.

 

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Last modified: 05/29/09