GSH(Glutathione) ELISA Kit

Alternative Names: L-Reduced Glutathione
Catalogue No. 7846-ELK
Size 96T
Reactivity General
Range 1.57-100 ng/mL
Sensitivity 0.48 ng/mL

Description

Alternative Names: L-Reduced Glutathione
Catalogue No. 7846-ELK
Size 96T
Reactivity General
Range 1.57-100 ng/mL
Sensitivity 0.48 ng/mL
Assay Type Competitive Inhibition
Sample Type serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length 2h
Research Area Metabolic pathway;
Test principle This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with Glutathione(GSH) protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Glutathione(GSH). Next,Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Glutathione(GSH) in the samples is then determined by comparing the OD of the samples to the standard curve.
Standard Curve

Concentration (ng/mL) OD Corrected OD
100.00 0.212
50.00 0.357
25.00 0.567
12.50 0.895
6.25 1.139
3.13 1.585
1.57 1.624
0.00 2.028
Precision Intra-assay Precision (Precision within an assay):CV%<8%

Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays):CV%<10%

Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.

Recovery Matrices listed below were spiked with certain level of recombinant GSH and the recovery rates were calculated by comparing the measured value to the expected amount of GSH in samples.

Matrix Recovery range Average
serum(n=5) 82-94% 88%
EDTA plasma(n=5) 81-97% 89%
Heparin plasma(n=5) 84-97% 90%
Linearity The linearity of the kit was assayed by testing samples spiked with appropriate concentration of GSH and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Matrix 1:2 1:4 1:8 1:16
serum(n=5) 79-96% 85-99% 95-104% 82-96%
EDTA plasma(n=5) 85-96% 87-101% 89-97% 85-94%
Heparin plasma(n=5) 98-105% 92-101% 88-102% 87-102%
Note For Research Use Only
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