Norman,
We still use silver staining when sensitivity is a consideration. We can
see down to 5 to 10ng BSA with the method by Blum. We have looked at Sypro
Ruby but did not notice a significant increase in sensitivity to justify the
increase in cost. We use two silver stain methods:
Blum, H., Beier, J., Gross, H.J., Electrophoresis, 1987, 8, 93-99.
Shevchenko, A., Wilm, M., Vorm, O., Mann, M., Anal. Chem., 1996, 68,
850-858.
Both colloidal (G250) and traditional coomassie (R250) should get you down
to the 0.1ug or maybe slightly lower levels if you use prolonged (i.e.
>overnight) staining methods. A good general reference for protein staining
is:
Allen, R.C., Budowle, B., Protein Staining and Identification Techniques,
The Biotechniques Series on Molecular Laboratory Methods, Number 3, 1999,
Eaton Publishing, Biotechniques Books Division
(ISBN 1-881299-08-2).
Best regards,
Lloyd W. Sumner, Ph.D.
Head, Biological Mass Spectrometry
The Samuel Roberts Noble Foundation
PO Box 2180
2510 Sam Noble Parkway
Ardmore, OK 73402
Voice 580-221-7392
Fax 580-221-7380
Email lwsumner@Noble.Org
-----Original Message-----
From: Norman Watts [mailto:WATTS@calvin.niams.nih.gov]
Sent: Wednesday, December 13, 2000 7:15 AM
To: Recipients of ABRF List
Subject: Misc, SPS-PAGE, sensitive protein stain needed
Hi all,
What is currently considered to be the most sensitive and
dependable stain for detecting protein bands on SDS-PAGE
gels? I expect ca. 0.05 ug/band.
norman
-------------------------------------------------------
Norman Watts, PhD.
Lab. of Structural Biology
Building 6, Room B2-34
6 Center Drive MSC2717
NIAMS, National Institutes of Health
Bethesda, MD 20892-2717
U.S.A.
E-mail: watts@calvin.niams.nih.gov
Phone: 301-402-3418
Fax: 301-480-7629
-------------------------------------------------------
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