Rat Anti-KLH IgG ELISA Kit with Controls
The Rat Anti-KLH IgG ELISA Kit with Controls is based on a solid phase enzyme-linked immunosorbent assay (ELISA). The assay uses KLH for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated anti-rat IgG antibodies for detection. Test serum or plasma samples are diluted and incubated in the microtiter wells for 45 minutes. The microtiter wells are subsequently washed, and HRP conjugate is added and incubated for 30 minutes. Anti-KLH IgG molecules are thus sandwiched between immobilized KLH and the detection antibody conjugate. The wells are then washed to remove unbound HRP-labeled antibodies, and TMB Reagent is added and incubated for 20 minutes at room temperature. This results in the development of a blue color. Color development is stopped by the addition of stop solution, changing the color to yellow, and optical density is measured spectrophotometrically at 450 nm. The concentration of anti-KLH IgG is proportional to the optical density of the test sample.