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Rat HIF2a(Hypoxia Inducible Factor 2 Alpha) ELISA Kit
ELK7037
Size： Price：
48 T $320.00
96 T $458.00
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Overview

Product name: Rat HIF2a(Hypoxia Inducible Factor 2 Alpha) ELISA Kit
Reactivity: Rat
Alternative Names: EPAS1; HLF; MOP2; PASD2; BHLHE73; Endothelial PAS Domain 1; HIF-1 Alpha-Like Factor; Basic-helix-loop-helix-PAS protein MOP2; Class E basic helix-loop-helix protein 73
Assay Type: Sandwich
Sensitivity: 31 pg/mL
Standard: 5000 pg/mL
Detection Range: 78.13-5000 pg/mL
Sample Type: tissue homogenates, cell lysates and other biological fluids
Assay Length: 3.5h
Research Area: Cardiovascular biology;
Uniprot ID: Q9JHS1
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 Rat HIF2a. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Rat HIF2a. 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 Rat HIF2a, 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 Rat HIF2a in the samples is then determined by comparing the OD of the samples to the standard curve.

Standard curve

Concentration (pg/mL) OD Corrected OD
5000.00 1.996 1.901
2500.00 1.387 1.292
1250.00 1.144 1.049
625.00 0.807 0.712
312.50 0.587 0.492
156.25 0.256 0.161
78.13 0.212 0.117
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 HIF2a and the recovery rates were calculated by comparing the measured value to the expected amount of HIF2a in samples.
Matrix Recovery range Average
serum(n=5) 87-99% 93%
EDTA plasma(n=5) 87-99% 93%
Heparin plasma(n=5) 80-95% 87%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of HIF2a 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) 81-94% 93-107% 93-102% 91-105%
EDTA plasma(n=5) 85-100% 84-99% 85-97% 88-102%
Heparin plasma(n=5) 87-102% 82-91% 86-93% 81-94%