Human Ephrin A5 ELISA Kit

Assay Genie supplies a sandwich ELISA, double antibody method for the quantitative detection of human Ephrin-A5, a protein implicated in cell signaling and neuronal development and used as a biomarker in neurobiology, cancer research and developmental biology. The kit is reactive with human samples and accepts serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Detection is colorimetric, with absorbance read at 450 nm after TMB substrate development.

Reported linearity (n=5) at 1:2, 1:4 and 1:8 dilutions is 86–105%, 87–103% and 89–105% in serum; 83–96%, 84–101% and 84–95% in EDTA plasma; and 82–95%, 87–101% and 82–94% in heparin plasma. Mean recovery is 96% in serum (range 88–104%), 95% in EDTA plasma (range 88–99%) and 95% in heparin plasma (range 89–101%). The associated Uniprot entry is P52803, and the kit is listed under the developmental biology research area.

The supplied procedure is a sample protocol specific to each lot: reagents and TMB substrate are equilibrated to room temperature; standards, samples and a zero control are loaded onto the pre-coated plate and incubated at 37°C for 90 minutes; a biotin-labelled detection antibody is added and incubated for 60 minutes at 37°C; HRP-Streptavidin (SABC) is added for 30 minutes at 37°C; TMB substrate is developed in the dark for 10–20 minutes; and stop solution is added before absorbance is read at 450 nm. The kit is stored at 2–8°C for 12 months. Sample preparation notes describe serum (clot, centrifuge at 1000 × g for 20 minutes), plasma (anticoagulant tube, centrifuge at 1000 × g for 15 minutes at 2–8°C), tissue homogenate, cell culture supernatant (centrifuge at 2500 rpm for 5 minutes) and cell lysate workflows.