Cat. No.
MAK-0025
Product Name
Monkey Anti-KLH IgG1
Storage
4°C
Species
Monkey
Instructions
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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 anti-KLH (keyhole limpet hemocyanin) antibody levels allows easy assessment of immune system regulation. Animals are immunized with KLH while undergoing drug treatment, and serum is collected at appropriate times post immunization. Typically, serum collected 5-7 days after immunization is used for measurement of anti-KLH IgM levels, and serum collected 14+ days post immunization is used to measure anti-KLH IgG levels. Comparison of anti-KLH IgG or IgM levels in drug treated versus control groups reveals effects of the drug on the immune response. This test kit allows rapid and quantitative measurement of anti-KLH IgG1 levels in serum or plasma. IgG1 is the major IgG subclass in monkeys.
Principle Of The Assay
The monkey anti-KLH IgG test kit is a solid phase enzyme-linked immunosorbent assay (ELISA). It uses KLH for solid phase (microtiter wells) immobilization and a horseradish peroxidase (HRP) conjugated mouse monoclonal anti-monkey IgG1 antibody for detection.A 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-KLH IgG1 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 IgG1 is proportional to the optical density.
