1. Chmiluminescence detection step is quite weird. You have to put membrane in between two plastic covers. However, if you put multiple membranes it may give higher background than single membrane-development.2. Blocking buffer may not sufficient if your samples need to be highly diluted.3. All the protocol is considered for one membrane blot; therefore, if you are analyzing multiple membrane, you may have trouble in unequal volume. (If you make 4 mL buffer, you will never get 4 mL.)4. I wish there is a tool for getting signal intensity. Software does not provide intensity-quantification function.

Review by JHA, on Feb 15, 2018