Cat. No.
MAK-0037
Product Name
Chicken Anti-SRBC IgM
Storage
2-8℃
Species
Chicken
Instructions
The assay was designed for use with serum or plasma obtained from chickens 5-7 days after immunization with SRBC.
Introduction
Measurement of anti-SRBC (sheep red blood cell) immunoglobulin levels is used to assess immune function in chickens. To date, a somewhat qualitative hemagglutination assay has often been used to measure antibody titers. Disadvantageously, the hemagglutination assay does not readily differentiate between IgM and IgG (IgY) responses. To address these issues, we at Life Diagnostics, Inc. have developed an enzyme linked immunosorbant assay (ELISA) that allows rapid and quantitative measurement of chicken anti-SRBC IgM levels in serum or plasma. We also manufacture a companion ELISA (catalog number SRBCG-5) that allows quantitative measurement of chicken anti-SRBC IgG levels. Studies at Life Diagnostics, Inc. have demonstrated that IV immunization of chickens with 0.5 ml of a 10% solution of SRBC caused an elevation of anti-SRBC IgM levels from ~200 units/ml on day zero to ~11,000 units/ml on day seven.
Principle Of The Assay
The chicken anti-SRBC IgM ELISA uses detergent solubilized SRBC ghosts for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated anti-chicken 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 and is derived from a standard curve.
