Gel storage before immunoblotting
clarosa at biocomp.unl.edu
Sat Apr 28 09:21:47 EST 2001
And you can transfer the bands to matrix for immunoblotting?? How
sensitive is the staining compared to coomassie?
On Fri, 27 Apr 2001, Dr. Hiranya S. Roychowdhury wrote:
> I will have to dig up the reference. I do it routinely. Very simple:
> 1. Rinse the gel, with dist. water, after SDS-PAGE.
> 2. Soak the gel in 0.3 to 0.4M CuCl2 (made in dist. water). The stain may
> be reused several times.
> 3. View/photograph the gel with incandescent lighting, incident at an
> angle. The exposure timing is to be worked out (usually 1/120th of a sec 16f).
> Yes, the gel can be stored, either in plain (dist.) water or in the CuCl2
> soln. at 4 C for several weeks, if not months. I have not noticed any loss
> of resolution or of polypeptide bands after two mnths of storage in CuCl2
> soln. or in water.
> Destaining is done with 0.25M EDTA in Tris.Cl, pH 9 (100mM), following
> which the staining is also lost.
> At 10:28 AM 4/27/01 -0500, clarosa wrote:
> >PLease give reference on CU-stain...
> >"Dr. Hiranya S. Roychowdhury" wrote:
> >> With Cu-stain, the gel may be stored for several weeks before transfer
> >> At 10:21 PM 4/11/01 +0100, Choe Woo-seok wrote:
> >> >After SDS-PAGE, can the gel be stored before electroblotting or is it
> >> >better to
> >> >be transfered immediately.
> >> >In
> Dr. Hiranya Sankar Roychowdhury
> College Asst. Prof.
> Molecular Biology,
> Dept. of Chemistry & Biochemistry
> Box 30001 - 3MLS
> New Mexico State University
> Las Cruces, NM 88003
> Lab: (505) 646 4722
> Office: (505) 646 8256
> hroychow at nmsu.edu
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