Cat. No.
MAK-0033
Product Name
Mouse Anti-SRBC IgM
Storage
4°C
Species
Mouse
Instructions
The assay was designed for use with serum or plasma obtained from mice five days after immunization with SRBC, at which point the immune response originates almost exclusively from IgM.
Introduction
Recent studies have demonstrated that suppression of anti-SRBC (sheep red blood cell) IgM levels by therapeutic agents serves as a useful indicator of immunosupression. This test kit allows rapid and quantitative measurement of mouse anti-SRBC IgM levels in serum or plasma samples.
Principle Of The Assay
The mouse anti-SRBC IgM test kit is based on a solid phase enzyme-linked immunosorbent assay (ELISA). The assay uses detergent solubilized SRBC ghosts2 for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated antimouse 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-SRBC IgM molecules are thus sandwiched between immobilized SRBC antigens 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-SRBC IgM is proportional to the optical density
of the test sample.
