Mouse Macrophage Inflammatory Protein 1 Alpha ELISA Kit

Sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, intended for the in vitro quantitative determination of mouse MIP-1 alpha/CCL3 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. An antibody specific for MIP-1 alpha/CCL3 is pre-coated onto the microplate. Standards and test samples are added to the wells, the captured analyte is bound by a Biotinylated Anti-MIP-1 alpha/CCL3 Antibody, and an HRP-Avidin conjugate is then added. TMB substrate is used to visualise the HRP enzymatic reaction, producing a blue-coloured product that changes to yellow after addition of an acidic stop solution. The density of yellow is proportional to the amount of analyte captured, and the concentration is calculated from the O.D. absorbance at 450 nm read on a colorimetric microplate reader.

The quantitative dynamic range is 15.6-1000 pg/ml, with a sensitivity of 5.8 pg/ml. Spike-and-recovery has been validated in serum (range 83-100%, average 91%), EDTA plasma (90-103%, average 96%) and heparin plasma (89-102%, average 95%) with n=5 per matrix. Dilution linearity has been characterised across 1:2, 1:4, 1:8 and 1:16 dilutions (n=5 per matrix), with serum returning 83-110%, 79-100%, 90-98% and 82-108%; EDTA plasma returning 90-98%, 90-105%, 73-109% and 87-91%; and heparin plasma returning 88-103%, 77-105%, 95-98% and 87-110% across those four dilutions respectively. Reactivity is confirmed for mouse MIP-1 alpha/CCL3.

Total assay time is 4 hours 30 minutes. Kit components are stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.