Cat. No.
MAK-0022
Product Name
Mouse Anti-KLH IgM
Storage
4°C
Species
Mouse
Instructions
This kit is designed to measure anti-KLH IgM levels in serum collected 5 days after immunization with KLH.
Introduction
Measurement of KLH (keyhole limpet hemocyanin) induced antiKLH antibody levels allows quantitative evaluation of the immune response. This ELISA is designed for the rapid and quantitative measurement of mouse anti-KLH IgM levels in mouse serum or plasma. A companion ELISA, catalog number KLHG-1, can be used for measurement of mouse anti-KLH IgG.
Principle Of The Assay
The mouse 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-mouse IgM antibodies for detection. Test serum or plasma samples are diluted and incubated in the microtiter wells for 1 hour. 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. Optical density is measured spectrophotometrically at 450 nm. The concentration of anti-KLH IgM is proportional to the optical density of the test sample.
