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Human MTHFR(Methylenetetrahydrofolate Reductase) ELISA Kit
ELK4174
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48 T $320.00
96 T $458.00
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Overview

Product name: Human MTHFR(Methylenetetrahydrofolate Reductase) ELISA Kit
Reactivity: Human
Alternative Names: 5,10-Methylenetetrahydrofolate Reductase(NADPH)
Assay Type: Sandwich
Sensitivity: 22.9 pg/mL
Standard: 4000 pg/mL
Detection Range: 62.5-4000 pg/mL
Sample Type: serum, plasma, tissue homogenates and other biological fluids
Assay Length: 3.5h
Research Area: Enzyme & Kinase;
Uniprot ID: P42898
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 MTHFR. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human MTHFR. 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 MTHFR, 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 MTHFR 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
4000.00 1.990 1.905
2000.00 1.633 1.548
1000.00 1.067 0.982
500.00 0.847 0.762
250.00 0.548 0.463
125.00 0.323 0.238
62.50 0.215 0.130
0.00 0.085 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 MTHFR and the recovery rates were calculated by comparing the measured value to the expected amount of MTHFR in samples.
Matrix Recovery range Average
serum(n=5) 79-93% 86%
EDTA plasma(n=5) 82-95% 88%
Heparin plasma(n=5) 85-97% 91%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of MTHFR 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) 84-99% 83-95% 83-95% 96-101%
EDTA plasma(n=5) 85-100% 84-98% 97-101% 88-96%
Heparin plasma(n=5) 86-93% 91-96% 87-98% 95-102%