ELISPOT for IFN-gamma and IL-10
时间:2005-07-18 00:00 来源:Scienceboard.net 作者:admin 点击:次
| Wednesday, October 29, 2003
Description An ELIspot assay to count cytokine-producting cells Procedure - Coat plates with primary Antibody 2 g/ml, 50 l/well - Incubate overnight 4C - Aspirate (do not remove the cover to keep the plate sterile) - Incubate with antigen and cells (vol. 200 l) overnight 37C - Wash 3 times like ELISA + 2 washings by aspiration - Add secondary antibody (biotinylated) 1 g/ml, 100 l/well - Incubate 45 minutes room Temp. - Wash 3 times (aspiration) - Add Avidin-peroxydase 1 g/ml, 100 l/well - Incubate 30 minutes room Temp. - Wash 5 times (aspiration) - Add substrate 41CN (sigma), allow coloration to develop - Wash 5 times with distilled water (aspiration) - Count spots Recipes Supplies For this assay, the Abs and plates were obtained from R&D systems Tips (责任编辑：泉水) |