Cat. No.
MAK-0036
Product Name
Chicken Anti-SRBC IgG
Storage
4°C
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 IgG (IgY) and IgM 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 IgG levels in serum or plasma. We also manufacture a companion ELISA (catalog number SRBCM-5) that allows quantitative measurement of chicken anti-SRBC IgM 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 IgG levels from undetectable levels on day zero to approximately 11,000 units/ml on day seven.
Principle Of The Assay
The chicken anti-SRBC IgG ELISA uses detergent solubilized SRBC ghosts for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated anti-chicken IgG antibodies for detection. Serum or plasma samples are diluted and incubated in the microtiter wells for 45 minutes. The microtiter wells are subsequently washed. HRP conjugate is added and incubated for 45 minutes. If Anti-SRBC IgG molecules are present, they are sandwiched between immobilized SRBC antigens 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 anti-SRBC IgG is proportional to the optical density of the test sample and is derived from a standard curve.
