Human Macrophage Migration Inhibitory Factor ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com is designed for the in vitro quantitative determination of human Macrophage Migration Inhibitory Factor (MIF) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.

The assay uses a pre-coated microplate arranged as 12 x 8 well strips and follows a sandwich immunoassay format. Standards and samples are added to wells, followed by a Biotinylated Anti-MIF Antibody, an HRP-Avidin conjugate, and a TMB substrate solution; catalysis produces a blue-coloured product that turns yellow after addition of acidic stop solution. The intensity of yellow, read as O.D. absorbance at 450 nm on a colorimetric microplate reader, is proportional to the amount of MIF captured and is calculated against a standard curve. The stated range is 0.781-50 ng/ml with a sensitivity of <0.29 ng/ml, and the total assay time is 4h 30m.

Spike-and-recovery performance is reported for serum (average 91%, range 80-102%, n=5), EDTA plasma (average 90%, range 81-100%, n=5) and heparin plasma (average 84%, range 80-89%, n=5). Dilution linearity across 1:2, 1:4, 1:8 and 1:16 is reported for serum (87-107%), EDTA plasma (84-108%) and heparin plasma (82-105%). Kit components include the pre-coated plate, lyophilized standard, Detection Solution A, Detection Solution B, 30X Wash Buffer, Sample Dilution Buffer, TMB Substrate, Stop Solution and plate sealers, stored at +4°C or -20°C as instructed.