Cat. No.
MAK-0039
Product Name
Rat Anti-Tetanus Toxoid IgM
Storage
2-8°C
Species
Rat
Instructions
The assay was designed for use with serum or plasma obtained from rats 5 days after immunization with tetanus toxoid, at which point the immune response originates predominantly from IgM.
Introduction
Evaluation of the levels of anti-tetanus toxoid IgM after immunization with tetanus toxoid provides a useful indicator of aspects of the immune response. The rat anti-tetanus toxoid IgM test kit developed by Life Diagnostics, Inc. facilitates rapid and quantitative measurement of rat anti-tetanus toxoid IgM levels in serum or plasma samples.
Principle Of The Assay
The rat anti-tetanus toxoid IgM test kit is based on a solid phase enzyme-linked immunosorbent assay (ELISA). The assay uses tetanus toxoid for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated anti-rat IgM antibodies for detection. Standards and diluted serum or plasma samples are 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-tetanus toxoid IgM molecules are thus
sandwiched between immobilized tetanus toxoid 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-tetanus toxoid IgM is proportional to the
optical density. Anti-tetanus toxoid IgM levels in the samples are derived by reference to a standard curve.
