Cat. No.
MAK-0090
Product Name
Rat IgG1
Storage
4°C
Species
Rat
Instructions
Test samples are diluted and incubated in the anti-rat IgG1 coated microtiter wells for 45 minutes alongside rat IgG1 standards.
Introduction
Four subclasses of IgG are present in rat: IgG1, IgG2a, IgG2b and IgG2c. Respective concentrations in 80-day old Lewis rats were found to be 0.10, 0.95, 2.06 and 0.09 mg/ml . Levels of the different subclasses vary with age and in response to immune stimulus. The rat IgG1 ELISA kit is designed for measurement of IgG1 in rat serum or plasma. The assay uses mouse monoclonal anti-rat IgG1 for solid phase (microtiter wells) immobilization and horseradish peroxidase (HRP) conjugated mouse monoclonal anti-rat IgG1 antibodies for detection. When used as directed, the kit recognizes only IgG1 in rat serum. It does not recognize mouse IgG. Crossreactivity with immunoglobulins from other species has not been investigated.
Principle Of The Assay
Test samples are diluted and incubated in the anti-rat IgG1 coated microtiter wells for 45 minutes alongside rat IgG1 standards. The microtiter wells are subsequently washed, and anti-rat IgG-HRP conjugate is added and incubated for 45 minutes. IgG1 molecules are thus sandwiched between the immobilization and detection antibodies. The wells are then washed to remove unbound HRPlabeled 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 IgG1 is proportional to the optical density of the test sample and is derived from a standard curve.
