Cat. No.
MAK-0041
Product Name
Monkey Anti-Tetanus Toxoid IgM
Storage
4°C
Species
Monkey
Instructions
The assay was designed for use with serum or plasma obtained from monkeys 5 days after immunization with tetanus toxoid, at which point the immune response originates predominantly from IgM.
Introduction
Drug candidates are routinely screened for evidence of immune system regulation during the discovery process. It is important that the immune response is not enhanced or diminished since such effects may lead to hypersensitivity or increased susceptibility to infection. Determination of a drug candidate’s effects on antitetanus toxoid antibody levels allows easy assessment of immune system regulation. Animals are immunized with tetanus toxoid while undergoing drug treatment and serum is collected at appropriate times post immunization. Serum collected 5-7 days after immunization is used for measurement of anti-tetanus toxoid IgM levels, and serum collected 14+ days post immunization is used to measure anti-tetanus toxoid IgG levels. Comparison of anti-tetanus toxoid IgG or IgM levels in drug treated versus control groups reveals effects on the immune response. This test kit allows rapid and quantitative measurement of antitetanus toxoid IgM levels in rhesus or cynomolgus monkey serum or plasma.
Principle Of The Assay
The monkey anti-tetanus toxoid IgM test kit is based on a solid phase enzyme-linked immunosorbent assay (ELISA). The assay uses tetanus toxoid1 for solid phase (microtiter wells) immobilization and a horseradish peroxidase (HRP) conjugated anti-monkey IgM monoclonal antibody (LDI clone 2C11-1-5) for detection. Standards and diluted serum or plasma samples are incubated in the microtiter wells for 60 minutes. 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. 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 antitetanus toxoid IgM is proportional to the optical density. Anti-tetanus toxoid IgM levels in the samples are derived by reference to a standard curve.
