Has anybody tried to fixate proteins on membrane (nitrocellulose or PVDF) by treatment with glutaraldehyd?
Somebody told me to do so to avoid protein loss from stripping?

If yes, could you give me the protocol or recipe please?

This is something I saw in a paper: 0.1% glutaraldehyde in PBS with 0.05% Tween 20

The membrane was fixed for 10 mins and then rinse with PBST. Then blocked with 5% milk in PBST, primary antibody incubation....

I haven't tried this myself yet. They don't say but it seems that they were able to strip the blot and reprobe for other proteins.

Here is the ref: Kristan et al (2006) Isolation of Mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation. J of Neuro Methods, 152, pp 136-143

JP

I never crosslinked my proteins, and I'm stripping in tris HCl pH6.8 50 mM, SDS 2% beta-mercaptoethanol 100mM, 30 minutes at 50°C. I don't loose so many proteins. I always start blotting with the lowest affinity antibody, strip, and blot with the highest affinity antibody.

I know someone do western without transforming protein to membrane. Is there someone konw this IN GEL western and how to do that?