Storage of SDS/PAGE gels - a question
hroychow at NMSU.EDU
Mon May 8 17:33:48 EST 1995
On 7 May 1995 troianovsk_s at MSDISK.WUSTL.EDU wrote:
> >Is it possible to store SDS/PAGE gels after running them? My advisor
> >is curious as to whether or not you can do it without significan diffusion
> >of the proteins. Is it possible to do so for 2-3 days? If so, what are
> >the conditions? Finally, if you have a method for storage, is it amenable
> >to blotting following 2-3 days of storage?
> What I would say, that any treatment of proteins while they are within the
> gel matrix would fix them to acrylamide. By this way you actualy prevent
> protein diffusion. As soon as you intend to prevent diffusion of your
> proteins, you`ll fix them to matrix, so the future transfer (blotting) to
> membrane will grately reduced.
> I was told that there are some reversable fixation techniques which allow
> fix the gel (proteins to the matrix) and late wash fixage away to free the
> protein. I never used that.
> I was also told that there is some way to disolve acrylamide itself. It
> usualy used when it is expected to transfer (blotting) very high MW
> proteins to increase the transfer efficiancy. I never used that either.
> Nevertheless, I think that if you`ll fix the proteins to acrylamide to
> prevent diffusion, then you should be able to disolve acrylamide itself to
> free the proteins for transfer. Such an approach would solve your problem.
> Hope this helps>
I am not sure how the above comments by the obviously well meaning person
helps. Here is something concrete:
Yes, SDS-PAGE gels can be stored indefinitely.
The method of storage depends on the end use.
If the gel is to be stored only for future reference, it can be stained
and dried on a 3MM paper. The staining prior to drying has to follow
acetic acid-methanol staining - ie. CBB-R method.
Copper stained gels can be stored in water almost indefinitely without
any loss of resolution. I have stored Cu-stained gels for up to a month,
but that is by no means the maximum length of time I am told. The
advantage of Cu-staining lies in the fact that the proteins can be eluted
with almost 100% efficiency or the gel can be stained by other
conventional methods following destaining in a simple Tris/glycine buffer
or Tris/EDTA buffer. Cu-stained gels can be blotted easily since the
fixing is absolutely reversible.
If interested, contact me for further details.
Hiranya S. Roychowdhury
Plant Genetic Engineering Lab.
Box 3GL, NM State Univ.
Las Cruces, NM 88003
Phone: (505) 646-5785
hroychow at nmsu.edu
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