Using human serum as first antibody in Western/ELISA
Posted 28 June 2012 - 01:37 PM
Because we could not locate the specific antibody against our HAV epitope, we use HAV vaccinated patient serum as first antibody in Western and indirect ELISA. However the background of negative control, which only applying serum(first Ab)+2nd Ab (anti-human-AP conjugated) was too strong. Tried different blocking with 5%Milk, 3% BSA, and 0.5%TBST, and did not work. After blocking also tried to block with horse serum or FBS, and both did not work. Anyone has any suggestion? Your suggestion will be appreciated.
Posted 28 June 2012 - 01:46 PM
Posted 28 June 2012 - 04:26 PM
Posted 29 June 2012 - 11:27 PM