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Human CH25H(Cholesterol-25-Hydroxylase) ELISA Kit
ELK3800
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48 T $320.00
96 T $458.00
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Overview

Product name: Human CH25H(Cholesterol-25-Hydroxylase) ELISA Kit
Reactivity: Human
Alternative Names: h25OH; Cholesterol 25-monooxygenase
Assay Type: Sandwich
Sensitivity: 51 pg/mL
Standard: 8000 pg/mL
Detection Range: 125-8000 pg/mL
Sample Type: Serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;
Uniprot ID: O95992
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 Human CH25H. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human CH25H. 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 Human CH25H, 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 Human CH25H 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
8000.00 1.948 1.861
4000.00 1.527 1.440
2000.00 1.142 1.055
1000.00 0.927 0.840
500.00 0.556 0.469
250.00 0.426 0.339
125.00 0.134 0.047
0.00 0.087 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 CH25H and the recovery rates were calculated by comparing the measured value to the expected amount of CH25H in samples.
Matrix Recovery range Average
serum(n=5) 82-95% 88%
EDTA plasma(n=5) 83-98% 91%
Heparin plasma(n=5) 91-99% 95%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of CH25H 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) 89-99% 83-101% 93-104% 96-101%
EDTA plasma(n=5) 86-96% 93-104% 89-97% 85-94%
Heparin plasma(n=5) 86-93% 91-105% 87-98% 99-102%