AIF separation

3 posts / 0 new
Last post
hima12
hima12's picture
AIF separation

Hello,
I am using an AIF antibody from cell signalling technologies (#4642) by western blot analysis.
The main problem which I am finding is separating the precursor protein (67Kda) from the mature form (57Kda).

I have used both 10% as well as 7.5% gel and went down to the protein concentration of 10 micrograms, but still I am having problems getting distinct bands.

Any suggestions would be appreciated.
Thanks

heehawmcduff
heehawmcduff's picture
Hi there,

Hi there,
I've actually used this antibody as well :) but don't have the results in front of me at the moment.  One thing that I can tell you is that we used 12% gels when we were running westerns with it and we did get two distinct bands after subcellular fractionation (looking at the cytosolic and mitochondrial fractions).  I also remember that the antibody was pretty effective so we had to dilute it quite a lot.

hima12
hima12's picture
Hello Heehawmcduff,

Hello Heehawmcduff,
Thanks for the info. I will try it out and let you know how it worked out.
Hima