TMB Prestained Red ELISA Peroxidase Substrate  

TMB Prestained Red ELISA Peroxidase Substrate

Rockland Immunochemicals produces a wide variety of buffers and substrates for use in ELISAs. Antigen was diluted in ELISA Microwell Coating Stabilizer (p/n MB-063-0100) added to the microwell plate and incubated overnight at 4°C. The plate was then blocked with ELISA Microwell Blocking Buffer with Stabilizer (p/n MB-064-1000) for 2 hours. The primary antibody was diluted in PBS Fish Gel Concentrate (1:10)(p/n MB-066-0100), added to the plate, and allowed to incubate 1 hour at room temperature. HRP conjugated secondary antibody was diluted in HRP Conjugate Stabilizer (p/n MB-060-0100), added to the plate, and allowed to incubate for 30 minutes at room temperature. TMB Prestained Red ELISA Peroxidase Substrate (p/n TMBE-RED-0100) (Rows A-D) and TMB ELISA Peroxidase Substrate (p/n TMBE-1000) (rows E-H) were added to the plate and allowed to incubate for 30 minutes at room temperature. The reaction was then stopped with 1M HCl and read at 450nm. The Red TMBE has comparable background to Rockland's regular TMBE.