Hi,

From a long time I have been trying to get pure cytoplasmic and nuclear protein extracts from virus infected A549 cell lines. I have tried to check for the pure protein extracts of cytoplasm and nucleus with western blotting validation using TBP and beta-actin antibodies.

But everytime, I get bands in both cytoplasmic and nuclear protein samples (for both antibodies).

Please suggest/answer for the following two points:

1-) relevant antibody to check the purity as I need nuclear extracts free from cytoplasmic extracts.

2-) Also, the tested and validated lysis buffer (with composition if needs to be prepared in lab) to extracts pure cytoplasmic and nuclear protein extracts.

Will be grateful for the expert opinions.

Thanks