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ELK Biotechnology

ELK9679

Rabbit GAPDH(Glyceraldehyde-3-Phosphate Dehydrogenase) ELISA Kit

Rabbit GAPDH(Glyceraldehyde-3-Phosphate Dehydrogenase) ELISA Kit

Sensitivity

Rabbit GAPDH(Glyceraldehyde-3-Phosphate Dehydrogenase) ELISA Kit

Overview

Product name: Rabbit GAPDH(Glyceraldehyde-3-Phosphate Dehydrogenase) ELISA Kit
Reactivity: Rabbit
Alternative Names: G3PD; GAPD; Peptidyl-cysteine S-nitrosylase GAPDH
Assay Type: Sandwich
Sensitivity: 0.56 ng/mL
Standard: 100 ng/mL
Detection Range: 1.57-100 ng/mL
Sample Type: tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;Metabolic pathway;Cardiovascular biology;Hepatology;Nutrition metabolism;
Test principle: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Rabbit GAPDH. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rabbit GAPDH. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Rabbit GAPDH, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. 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 Rabbit GAPDH 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 2.126 2.031
50.00 1.673 1.578
25.00 1.273 1.178
12.50 0.892 0.797
6.25 0.525 0.430
3.13 0.327 0.232
1.57 0.241 0.146
0.00 0.095 0.000

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 GAPDH and the recovery rates were calculated by comparing the measured value to the expected amount of GAPDH in samples.
Matrix Recovery range Average
serum(n=5) 79-93% 86%
EDTA plasma(n=5) 80-95% 87%
Heparin plasma(n=5) 93-107% 100%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of GAPDH 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) 83-89% 88-104% 97-103% 89-101%
EDTA plasma(n=5) 87-98% 92-104% 85-96% 86-95%
Heparin plasma(n=5) 90-102% 81-90% 87-95% 83-96%
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