Principle of Assay
Human ATPase, Na+/K+ Transporting Beta 4 ELISA Kit (A7022) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human ATPase, Na+/K+ Transporting Beta 4 in tissue homogenates, cell lysates or other biological fluids. An antibody specific for ATPase, Na+/K+ Transporting Beta 4 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the ATPase, Na+/K+ Transporting Beta 4 present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-ATPase, Na+/K+ Transporting Beta 4 Antibody, which binds the captured ATPase, Na+/K+ Transporting Beta 4 present in each well. HRP-Avidin conjugate is then added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of ATPase, Na+/K+ Transporting Beta 4 captured in each well. The concentration of ATPase, Na+/K+ Transporting Beta 4 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.