Human Ceramide kinase (CERK) ELISA Kit

Designed for the quantitative measurement of Human Ceramide kinase (CERK), this Sandwich ELISA from Assay Genie employs a colorimetric, double-antibody detection method. The assay is reactive with human samples including serum, plasma, tissue homogenates, cell culture supernates and other biological fluids. Reported performance includes a detection range of 0.625–40 ng/mL, a sensitivity of 0.31 ng/mL, an intra-assay CV of 4.7% and an inter-assay CV of 8.7%.

The microplate is pre-coated with a capture antibody specific to the target protein. Standards and appropriately diluted samples are added to the wells and incubated to allow the analyte to bind. After washing, a biotinylated detection antibody directed against a different epitope is added, forming a sandwich complex. HRP-conjugated Streptavidin is then introduced, followed by TMB substrate to generate a colorimetric reaction. The reaction is stopped with an acidic solution and absorbance is read at 450 nm using a microplate reader; signal intensity is directly proportional to analyte concentration and quantified against a standard curve.

Linearity has been evaluated at dilutions of 1:2, 1:4, 1:8 and 1:16 across serum, EDTA plasma and heparin plasma. Average recovery was 97% (range 91–103%) for serum and 99% (range 93–105%) for plasma. Ceramide kinase catalyzes the phosphorylation of ceramide to form ceramide 1-phosphate, acting efficiently on natural and analog ceramides (C6, C8, C16 ceramides and C8-dihydroceramide), to a lesser extent on C2-ceramide and C6-dihydroceramide, but not on other lipids such as sphingosines. The enzyme has been implicated in cellular processes including proliferation, apoptosis, phagocytosis and inflammation.