Cat. No.
MAK-0047
Product Name
Mouse Anti-TNP IgM
Storage
2-8℃
Species
Mouse
Instructions
This kit is designed to measure anti-TNP IgM levels in serum collected ~7 days after immunization with TNP-carrier protein conjugates
Introduction
Haptens such as trinitrophenol (TNP), when attached to carrier proteins such as ovalbumin or keyhole limpet hemocyanin (KLH) and injected into animals, produce a strong immune response. By measuring changes in the levels of anti-TNP IgM and IgG in appropriate animal models, researchers can assess the impact of pharamacologic or genetic manipulations on the immune system. Life Diagnostics, Inc. offers a simple ELISA for measurement of mouse anti-TNP IgM. We also manufacture an ELISA kit for mouse anti-TNP IgG.
Principle Of The Assay
The mouse anti-TNP IgM test kit is based on a solid phase enzymelinked immunosorbent assay (ELISA). The assay uses TNP-BSA as the capture antigen (coated on the microtiter wells) and horseradish peroxidase (HRP) conjugated anti-mouse IgM antibodies for detection. Serum or plasma samples are diluted and incubated alongside standards in the microtiter wells for 1 hour. The wells are subsequently washed, and HRP conjugate is added and incubated for 45 minutes. Anti-TNP IgM molecules are thus sandwiched between immobilized TNP 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-TNP IgM is proportional to the absorbance at 450 nm and is derived from a standard curve.
