Nephrat II is a direct competitive ELISA for the quantitative determination of urinary albumin in rat specimens. To complete the assay, controls, standards and diluted samples are added to rat albumin coated plates. An anti-rat albumin antibody-HRP conjugate is added, and allowed to react. Specific antigen-antibody reactions occur either with the albumin of the fluid phase or with that of the stationary phase, hence the notion of competitive binding. Subsequently, the plate is washed, and unbound reactants are washed away. Only conjugate bound to the stationary phase remains. A chromogenic substrate, TMB, is added to the wells, and color development ensues. Color intensity is inversely proportional to the log of rat albumin concentration of the sample.
Nephrat II has a useful dynamic range of 1.56-100 ug rat albumin/ mL, and samples must be diluted appropriately to fall within that range. The kit includes 2 plates and sufficient reagents to complete 80 analyses in duplicate.