Changes

#Block with 2% BSA in [[ TBST ]] or 5% skim milk powder in TBST for >1h

#Incubate with primary antibody (check for dilution) in 1 mg/mL BSA for >1h

#Incubate with appropriate secondary antibody (10 000X) for 45min-1h

#Rinse once or twice with double distilled water

#Prepare ECL by mixing one volume of the black bottle solution with one volume of the white bottle solution (be careful not to cross-contaminate ECL bottles with wrong solution).