Colloidal Gold total protein stain

Shaun D. Black SHAUN at JASON.UTHCT.EDU
Wed Nov 1 11:19:57 EST 1995


Hi all,

     LOGAND asked about total protein stains for western blots.  Here's the
protocol for our lab's procedure:

1. Prepare colloidal gold staining solution (see Yamaguchi, K., and
Asakawa, H. (1988) Preparation of colloidal gold for staining proteins
electrotransferred onto nitrocellulose membranes.  Anal. Biochem. 172,
104-107) by adding 5 mL 1% HAuCl4 (aq), 6 mL 35% formalin, and 5 mL Tween 20
to 480 mL pure water in sequence with rapid stirring; then add 3 mL 0.2 N KOH
dropwise.  After vigorous stirring overnight, the stain will have a deep
red color; adjust pH to 3.5 with formic acid and store in the dark at
4 degrees C.

2.  Soak blot in about 25 mL of colloidal gold reagent with gentle agitation
until bands are adequately developed/stained.  Destain in purified water.
Stained membranes can be dried at room temperature for storage.

It's a pretty straightforward procedure, even if you make your own reagents.
If you want to buy colloidal gold stain, we have used the Bio-Rad reagent
with success.

Best regards,  Shaun

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  = Shaun D. Black, PhD   | Internet address:     shaun at jason.uthct.edu = 
  = Dept. of Biochemistry | University of Texas Health Center, at Tyler = 
  = World Wide Web:    http://pegasus.uthct.edu/UTHCT-Home/Welcome.html =
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