Cat. No.
MAK-0024
Product Name
Rat Anti-KLH IgM
Storage
4°C
Species
Rat
Instructions
The assay was designed for use with serum or plasma obtained from rats five days after immunization with KLH, at which point the immune response originates almost exclusively from IgM.
Introduction
Recent studies have demonstrated that suppression of anti-KLH (keyhole limpet hemocyanin) IgM levels by therapeutic agents serves as a useful indicator of immunosupression. This ELISA allows rapid and quantitative measurement of rat anti-KLH IgM levels in serum or plasma samples.
Principle Of The Assay
The rat anti-KLH IgM test kit is based on a solid phase enzymelinked immunosorbent assay (ELISA). The assay uses KLH for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated anti-rat IgM 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 45 minutes. Anti-KLH IgM 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 IgM is proportional to the optical density of the test sample.
