RE: silver staining IEFgels

Lila Castellanos Serra (lila.castellanos@cigb.edu.cu)
Mon, 18 Oct 1999 09:00:38 -0400

Jaime
We have good results with the protocol from Heukeshoven and Dernick
(Electrophoresis 1985 vol 6 103-112)
1. Fix in Ethanol/Acetic acid (40%:10%) for 30 min.
2. Incubate in tiosulfate:sodium acetate:glutardialdehyde:etanol
(0.5g:10.3g:1.3 ml:75 ml, in water up to 250 ml) for 2h or longer
3.Wash with abundant water 3 x 20 min
4.Stain in AgNO3: formaldehyde (0.25g:50 ul in 250 ml water) for 45 min
5. Wash with water briefly (10 s)
6. Development in sodium carbonate:formaldehyde (6.25 g: 25 ul in 250 ml
water). Renew this solution after 1-2 min.
7. Stop development in EDTA (2.92 g in 200 ml water)

Alternatively, follow the procedure from Blum et al (Electrophoresis 1997,
vol 8, 93-99) compatible with MS analysis but slightly less sensitive. This
was reported for SDS gels but it also works for IEF. (See also Arnot et al,
Anal Biochem 1998, vol 258, 1-18 for its application to SDS gels)

Regards
Lila Castellanos-Serra
CIGB
Havana
-----Original Message-----
From: jaime paba martinez [SMTP:bucaros@unb.br]
Sent: Friday, October 15, 1999 10:24 AM
To: Recipients of ABRF List
Subject: silver staining IEFgels

Dear All,
I would like to know if it is possible to silver stain ampholine
IEF
gels, not IPG. A good reference would be great.

Thanks in advance,

Jaime Paba
Universidade de Brasilia
Brazil