ELISA Kit for 5-Methyltetrahydrofolate (5-Me-THF)

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to 5-Me-THF has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled 5-Me-THF analogues and unlabeled antigen (Standards or samples) with the pre-coated antibody. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of 5-Me-THF in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of 5-Me-THF in the sample.

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level 5-Methyltetrahydrofolate (5-Me-THF) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level 5-Methyltetrahydrofolate (5-Me-THF) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV Inter-Assay: CV Inter-Assay: CV

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

This assay has high sensitivity and excellent specificity for detection of 5-Methyltetrahydrofolate (5-Me-THF) . No significant cross-reactivity or interference between 5-Methyltetrahydrofolate (5-Me-THF) and analogues was observed.