Wednesday, October 29, 2003
Description A simple ELISA assay to measure cytokines in culture supernatants Procedure - Coat plates with primary Antibody 2 g/ml, 50 l/well - Incubate overnight 4C - Wash 3 times - Block with 10% FCS, 200 ml/well - Incubate 2 hours room Temp. - Add standards and sample, 100 ml/well - Incubate 45 minutes room Temp. - Wash 3 times - Add secondary antibody (biotinylated) 1 g/ml, 100 l/well - Incubate 45 minutes room Temp. - Wash 3 times - Add Avidin-peroxydase 1 g/ml, 100 l/well - Incubate 30 minutes room Temp. - Wash 3 times - Add substrate ABTS (remember to thaw it and add 10 ml of H2O2 per 10 ml of substrate). - Read 405 nm Recipes Supplies Ab were purchased from R & D sytems ELISA plates were Dynatech Tips (责任编辑:泉水) |