Description
Introduction: Podocin is a protein which lines the podocytes and assists in maintaining the barrier at the glomerular basement membrane. Podocin is a new member of the stomatin family, which consists of hairpin-like integral membrane proteins with intracellular NH2- and COOH-termini. Mutations in the podocin gene NPHS2 can cause nephrotic syndrome, such as Focal Segmental Glomerulosclerosis (FSGS) or Minimal Change Disease (MCD).
Principle of the Assay: The microtiter plate provided in this kit has been pre-coated with an antibody specific to podocin. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for podocin and Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB (3,3',5,5' tetramethyl-benzidine) substrate solution is added to each well. Only those wells that contain podocin, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a 3 sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm +/- 2 nm. The concentration of podocin in the samples is then determined by comparing the O.D. of the samples to the standard curve