5-Methyltetrahydrofolate ELISA Kit

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<10% Inter-Assay: CV<12%

5-Methyltetrahydrofolate is a methylated derivate of tetrahydrofolate. It is generated by methylenetetrahydrofolate reductase from 5,10-methylenetetrahydrofolate and used to recycle homocysteine back to methionine by 5-methyltetrahydrofolate-homocysteine methyltransferases (also called methionine synthases) .

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.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery Range (%) Average (%) Serum (n=5) 84-99 92 EDTA plasma (n=5) 96-105 102 Heparin Plasma (n=5) 80-102 93 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 84-99 92 Serum (n=5) 84-99 92 EDTA plasma (n=5) 96-105 102 EDTA plasma (n=5) 96-105 102 Heparin Plasma (n=5) 80-102 93 Heparin Plasma (n=5) 80-102 93

Component Amount Anti-5-Methyltetrahydrofolate Microplate 96 Wells (12 x 8 Well Strips) 5-Methyltetrahydrofolate Lyophilized Standard 2 100X 5-Methyltetrahydrofolate Biotin Complex 120 uL 100X Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Biotin Complex Diluent 1 x 12 mL Avidin/HRP Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-5-Methyltetrahydrofolate Microplate 96 Wells (12 x 8 Well Strips) Anti-5-Methyltetrahydrofolate Microplate 96 Wells (12 x 8 Well Strips) 5-Methyltetrahydrofolate Lyophilized Standard 2 5-Methyltetrahydrofolate Lyophilized Standard 2 100X 5-Methyltetrahydrofolate Biotin Complex 120 uL 100X 5-Methyltetrahydrofolate Biotin Complex 120 uL 100X Avidin/HRP Conjugate 120 uL 100X Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Biotin Complex Diluent 1 x 12 mL Biotin Complex Diluent 1 x 12 mL Avidin/HRP Conjugate Diluent 1 x 12 mL Avidin/HRP Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL