Ive just made a tagged over-expression line of my gene and I want to identify any protein-protein interactions. Does anyone have a protocol I can use, the procedure in my lab is to use Brain ring gland complex samples for a western followed by mass spectrometry. The issue is most of the time it doesn't work and takes a long time to dissect the samples. I was wondering if anyone has a protocol using whole body larvae I can use?
Thank you!
Sounds like an IP-WB, immune-precipitation, followed by western blot. You do an initial enrichment using an antibody against one of your protein targets, enriched using mag-beads or agarose beads or similar. Then you run the enriched lysate on western blot (SDS-PAGE FTW), and probe for your SECOND protein. If your two proteins are within close proximity to each other, then you should find it using that method. A more detailed and specific protocol can be found online using the reagents you intend to use (mag separation, etc).
Craig Silver by any chance do you have a drosophila protein extraction protocol I can use? Thank you so much!
- Badges
- Science topic
- Similar topics
- Mathematical Sciences
- Graphs