Hi all

I detected one molecule for my paper.

However, my band is faint/negative band to show.

Please, give me the comment for detecting this protein ?

I wrote my protocol for WB

1. Protein Loading (60 ug) :

2. 10 % Gel : Hand Made

3. Electrophoresis : 30 min 90 V + 60 min 120 V

4. Wet Transfer : 16 hour 30 V in PVDF

5. Blocking : 1 hour in 5% Skim milk in PBST

6. Primary AB : 1:2500 in 5% Skim Milk

7. Wash : 10 mins * 4 times

8. Secondary AB : 1:15000 in 5% Skim Milk

9. Wash : 10 mins * 4 times

10. Detection : Lumigen ECL.