non-interfering stain for 2D westerns?

Mitchell Isaacs misaacs at spam_me_not.student.usyd.edu.au
Wed Nov 14 18:24:42 EST 2001


"Paul Muhlrad" <pmuhlrad at u.arizona.edu> wrote in message
news:B817FDD6.5E76%pmuhlrad at u.arizona.edu...
> I'm looking for a protein stain for 2D protein gels and/or filter
membranes
> that is compatible with subsequent immuno-probing and HRP/chemiluminescent
> detection (e.g. SuperSignal).  I need sensitivity on the same order as
> silver staining.  Thanks for any suggestions.

Try Sypro ruby (refs below) - I don't have any personal knowledge of
compatabiliy with your techniques (it is fine with MS) - but its sensitivity
is very good. I think the first ref may answer some compatability issues.
There are quite a few other papers in Electrophoresis in the last couple of
years that either use or discuss Sypro stains.

Cheers,
Mitchell

Berggren K, Steinberg TH, Lauber WM, Carroll JA, Lopez MF, Chernokalskaya E,
Zieske L, Diwu Z, Haugland RP, Patton WF. A luminescent ruthenium complex
for ultrasensitive detection of proteins immobilized on membrane supports.
Anal Biochem 1999 Dec 15;276(2):129-43.

Berggren K, Chernokalskaya E, Steinberg TH, Kemper C, Lopez MF, Diwu Z,
Haugland RP, Patton WF. Background-free, high sensitivity staining of
proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide
gels using a luminescent ruthenium complex. Electrophoresis 2000
Jul;21(12):2509-21

Lopez MF, Berggren K, Chernokalskaya E, Lazarev A, Robinson M, Patton WF. A
comparison of silver stain and SYPRO Ruby Protein Gel Stain with respect to
protein detection in two-dimensional gels and identification by peptide mass
profiling. Electrophoresis 2000 Nov;21(17):3673-83







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