ELISA Microwell Blocking Buffer with Stabilizer (Azide and Mercury Free)  

ELISA Microwell Blocking Buffer with Stabilizer (Azide and Mercury Free)

ELISA results of ELISA Microwell Blocking Buffer with Stabilizer (Azide and Mercury Free) (p/n MB-064). Each well was coated in duplicate with 1.0 μg of Mouse IgG Whole Molecule. Blocking Buffer (p/n MB-064, lot#35968) (Red), Blocking Buffer reference lot# (Green), and Blocking Buffer control lot# (Blue). The starting dilution of antibody was 5μg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using Rabbit Anti-Mouse IgG, Goat anti-Rabbit HRP 1:8500, and TMB 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 ELISA Peroxidase Substrate (p/n TMBE-1000) was 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.