anti-GFP antibody

Model Organism: Worms Methods & Reagents
1

Can anyone recommend a good anti-GFP antibody for Westerns of transgenic worms?

Any that work well of IP?

2

Many probably work well, but I really like Abcam Ab290 (rabbit polyclonal) for westerns (from worms lysates).

Strong antibody, and can be used at low dilutions (1/10 000 or so). IPs well too. Not sure how the price compares to other anti-GFPs.

Terry

3

Thanks, I’ll try it.

4

for IP with anti-GFP antibodies should I use protein A or protein G? 4C over-night or 2hrs at room-temp? which one works better? Thanks in advance.

5

I like Abcam Ab290 as well. It works great!

6

Can any of you give me an idea of how many worms-worth of protein extract you use per lane in order to see a decent GFP band? Of course the intensity of the signal will depend on how strongly and in how many cells the transgene is expressed, but I’m wondering what a good starting point would be for a broadly-expressed, reasonably bright GFP.

Thanks!
Doug

7

I guess 50 adult worms look reasonable if the GFP expression is “decent”