Human Neopterin ELISA Kit

A competitive ELISA from Assay Genie for the quantitative measurement of human neopterin. The microplate is pre-coated with antigen; standards and appropriately diluted samples are added to the wells and incubated, followed by a biotin-labeled detection reagent, HRP-conjugated streptavidin, and TMB substrate. The reaction is stopped with an acidic solution and absorbance is read at 450 nm using a microplate reader, with signal intensity directly proportional to analyte concentration and quantified against a standard curve.

Validated sample types include serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The kit should be stored sealed at 2-8°C and not cryopreserved.

Linearity (n=10) for serum is 85-104% at 1:2, 86-99% at 1:4, and 89-105% at 1:8; for EDTA plasma 82-97%, 82-99%, and 84-101%; and for heparin plasma 80-99%, 87-100%, and 83-100%. Recovery ranges (n=10) are 87-101% with an average of 94% in serum, 88-105% with an average of 97% in EDTA plasma, and 97-103% with an average of 99% in heparin plasma. The sample protocol adds 50 µL of standard or sample together with 50 µL of biotin-labeled antibody per well, incubates at 37°C for 45 minutes, washes the plate five times with one-minute soaks, develops with TMB in the dark for 10-20 minutes, and reads absorbance at 450 nm after stop solution.