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Mouse H2AFX(H2A Histone Family, Member X) ELISA Kit
ELK0328
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48 T $320.00
96 T $458.00
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Overview

Product name: Mouse H2AFX(H2A Histone Family, Member X) ELISA Kit
Reactivity: Mouse
Alternative Names: H2AX
Assay Type: Sandwich
Sensitivity: 11.6 pg/mL
Standard: 2000 pg/mL
Detection Range: 31.25-2000 pg/mL
Sample Type: Serum, plasma, tissue homogenates and other biological fluids.
Assay Length: 3.5h
Research Area: Metabolic pathway;
Uniprot ID: P27661
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 Mouse H2AFX. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Mouse H2AFX. 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 Mouse H2AFX, 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 Mouse H2AFX 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
2000.00 2.035 1.940
1000.00 1.700 1.605
500.00 1.174 1.079
250.00 0.883 0.788
125.00 0.521 0.426
62.50 0.366 0.271
31.25 0.179 0.084
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 H2AFX and the recovery rates were calculated by comparing the measured value to the expected amount of H2AFX in samples.
Matrix Recovery range Average
serum(n=5) 89-101% 95%
EDTA plasma(n=5) 82-95% 88%
Heparin plasma(n=5) 80-94% 87%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of H2AFX 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) 91-99% 83-89% 88-104% 97-103%
EDTA plasma(n=5) 83-94% 85-96% 92-103% 86-97%
Heparin plasma(n=5) 81-92% 92-103% 87-98% 90-101%