Has anyone used the Roche anti-GFP Antibody (Cat#11814460001) to detect sfGFP on a western blot? If not, could you please suggest an antibody that detects sfGFP with relatively high sensitivity by western blotting? Thanks so much for your help!
I strongly recommend these antibodies from Abcam (AB290 crude rabbit polyclonal and
AB6556 affinity purified rabbit polyclonal). They work amazingly well. You can generally start at 1:10, 000 and go up to 1:100, 000 depending on your expression. You can also re-use the antibody quite a few times. Just don't try it overnight. 45 mins to an hour is more than enough with this antibody.
Thank you very much for the information! It is very helpful to know this.
Dear Dale,
Could you tell me whether you have some IHC experience too with the ab6556?
Nora,
I haven't done immunohisto with these antibodies, but I have done immuno-EM with the affinity purified rb anitbody as well as IF with the mouse monoclonal one (http://www.abcam.com/gfp-antibody-lgb-1-ab291.html). I should say that they are made by my old lab, but I would swear by these antibodies. They made my grad studies so much easier. You can also find them here: www.eusera.com
I hope this helps.
Hi Jyoti,
Did you ever try the antibodies against sfGFP? I'm also looking for good antibodies against this version of GFP, since I plan to do westerns and IP.
Hi Juan,
Sorry for the very delayed response. I did not see your question until now. It turns out that the Roche anti-GFP antibody does detect sfGFP on a western blot.
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