I performed WB with 15 ug/10 ul protein sample for 1 lane
My expected band is indicated in red marker in the image.
But something wrong with my band, always.
Band is connected by strong dots between the bands.
It seems like that my band is moved into the gap between band!!!
Here is my running condition
- 10% SDS-PAGE
- 15-20 ug / 10 ul for 1 loading (I tried 15 and 20 ug)
- starting V 100 > final V 160
- running time 75 min
- PVDF transfer
- 375 mA 70 min
- 5% skim milk blocking 1 h
Please anybody help me to solve this problem..