anti-GFP primary antibody optimized for Western Blotting against embryo lysate

Model Organism: Worms Methods & Reagents
1

Hi,

I was wondering if anyone knew of a suitable GFP primary antibody optimized for use in Western blotting vs. embryo lysates. The ideal antibody should be both highly sensitive and specific. Thanks for the help! I really appreciate it.

2

I haven’t done any GFP blots in embryos, but the following have been reported:
anti-GFP (Roche): http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2064726/
α-GFP antibody (JL-8 1:1000 dilution, BD Biosciences): http://www.ncbi.nlm.nih.gov/pubmed/16338136

Alternatively, the Developmental Studies Hybridoma Bank has generated a number of monoclonals against GFP (http://dshb.biology.uiowa.edu/Antibody-Collections/GFP). DSHB-GFP-12A6 is supposedly ChIP grade (I haven’t tested) and works well on western blots of whole worm lysates (haven’t tested the embryo). Importantly, it’s nice and cheap. DSHB monoclonals are $32 per ml of supernatant and $50 per 0.1 ml of concentrated antibody. It’s definitely worth trying out. They also have good and cheap Myc, GST, His, MBP and tubulin monoclonal antibodies.

best,

Jordan

3

Thank you Jordan, I will be sure to look into the DSHB antibodies.