Canine Interleukin 5 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com designed for the in vitro quantitative determination of canine Interleukin 5 (IL-5) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. The procedure uses a 96-well microplate pre-coated with an anti-IL-5 capture antibody; standards and samples are added, IL-5 is captured by the immobilized antibody, and a biotinylated anti-IL-5 antibody followed by an HRP–avidin conjugate is used for detection. TMB substrate produces a blue product that turns yellow after addition of the acidic stop solution, and absorbance is read at 450 nm, with the IL-5 concentration calculated from a standard curve.

The assay has a measuring range of 15.6–1000 pg/ml, a sensitivity of 5.4 pg/ml, and a total assay time of 4 h 30 min. It is run on a pre-coated microplate (12 x 8 well strips) and read with a colorimetric microplate reader. Spike-and-recovery has been evaluated in serum (n=5, 80–102%, average 91%), EDTA plasma (n=5, 81–99%, average 90%) and heparin plasma (n=5, 80–89%, average 84%). Dilutional linearity has also been characterised in serum (n=5), EDTA plasma (n=5) and heparin plasma (n=5) at dilutions of 1:2, 1:4, 1:8 and 1:16.

Kit components are stored at +4 °C or −20 °C as instructed, the kit should not be used past its expiration date, and it is intended for use with a colorimetric microplate reader.