IPs with GFP antibody

Model Organism: Worms Methods & Reagents
1

Hello. Just wondering if anyone is currently having success doing co-IP using an anti-GFP antibody
for the pull down (and worm lysate from a GFP-fusion protein-expressing strain), and if you would
be willing to share your methods. Thanks.

2

I know offhand of a couple of published papers (1, 2) that used the mAb3E6 monoclonal anti-GFP antibody from QBiogen in immunoprecipitations; you could consult the papers’ Experimental Procedures or ask their authors.

3

Thanks. Both Figure 5 from Berdichevsky et al. 2006 and Supporting Figure 8 from Harrison et al. 2006 used mouse monoclonal anti-GFP antibody 3E6 from Qbiogene,
now MP Biomedicals. We have been trying to use Clontech mAb JL-8. Has anyone tried this Clontech antibody for IPs in worms?

4

A quick Google search finds at least one publication, from the Seydoux lab, that did immunoprecipitations using the Clontech antibody.

5

Thanks. Sorry for missing the obvious. Stitzel et al. 2007 Figure 3C.

6

It would be particularly interesting to know if anyone has made direct comparisons with commercial GFP antibodies for optimization?