I have recently started a project where I will be using BIFC in S.cerevisiae to look for protein protein interactions. Can anyone suggest if anti GFP can identify Vn/Vc (singly)tagged proteins on Western Blot? I do not have antibodies for the tagged proteins.
Hi Malini,
Most commercially available antibodies against GFP also recognize YFP. In this case it will depend on the position of the epitopes. If your GFP anybody is a polyclonal raised against a large peptide of the GFP (or whole molecule) then you would have high chances that it will recognize both "halves" fused to your Vc/Vn taged proteins by Western Blot. However to really know you will have to express these constructs and check what does your antibody recognizes.
Best,
Thank you so much! I just checked the antibody and it is the IgG fraction! So I think using the polyclonal GFP antibody should help. This was really helpful. I have not done westerns before so I was probably not thinking critically. You directed me on the right track! thank you very much.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Antibodies