Hi all

I'm doing Sandwich ELISA , I wanted to coat the antibody to the wells , So I have many questions:

1: Coating buffer ( which is more accurate)

bicarbonate -carbonate buffer: at 100mM or 50mM

2: Coating and blocking time (which is more accurate)

• overnight at 4C
• 2h at 25C
• 1.30 h at 37C

3: Blocking buffer for ELISA

PBS with 5% milk ( I heard is recommend for WB : more suitable for western blot )

PBS with 1% BSA

PBS with 3% BSA

PBS with 5% milk and 1% BSA

Thanks .