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5-Me-THF(5-Methyltetrahydrofolate) ELISA Kit
ELK8059
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48 T $265.00
96 T $376.00
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Overview

Product name: 5-Me-THF(5-Methyltetrahydrofolate) ELISA Kit
Reactivity: General
Alternative Names: 5-Me-THFA; 5-Me-H4FA; 5-Me-THF; 5-Me-H4F; Levomefolic Acid; Metafolin; 5-Methyltetrahydrofolic Acid; L-Methylfolate
Assay Type: Competitive Inhibition
Sensitivity: 0.43 ng/mL
Standard: 90 ng/mL
Detection Range: 1.41-90 ng/mL
Sample Type: serum, plasma and other biological fluids
Assay Length: 2h
Research Area: Hematology;Reproductive science;Nutrition metabolism;
Test principle: This assay employs the competitive inhibition enzyme immunoassay technique. The microtiter plate provided in this kit has been pre-coated with 5-Me-THF protein. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to 5-Me-THF. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added. 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 5-Me-THF 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
90.00 0.197
45.00 0.357
22.50 0.611
11.25 0.812
5.63 1.197
2.82 1.511
1.41 1.796
0.00 2.213

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 5-Me-THF and the recovery rates were calculated by comparing the measured value to the expected amount of 5-Me-THF in samples.
Matrix Recovery range Average
serum(n=5) 92-106% 99%
EDTA plasma(n=5) 77-95% 86%
Heparin plasma(n=5) 88-103% 91%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of 5-Me-THF 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) 88-104% 84-98% 92-103% 82-96%
EDTA plasma(n=5) 86-97% 83-96% 91-103% 96-105%
Heparin plasma(n=5) 88-103% 90-98% 92-106% 85-97%