Silver staining of protein gels
This protocol is adapted from Shevchenko et al., Analytical chemistry 68, p 850-858 (1996) and is especially suited for 1 mm gels.
- fix the gel
by either Coomassie-staining / destaining or by using the following
incubations:
1. 30 min in 50% methanol / 10% acetic acid
2. 15 min in 5% methanol / 1% acetic acid - wash 3 x 5 min with H2O
- incubate 90 sec. in thiosulfate solution (0.2 g/l Na2S2O3 x 5 H2O)
- rinse 3 x 30 sec. with H2O
- incubate 30 min with 0.2 g/100 ml AgNO3
- develop for up to 10 min in fresh thiosulfate developing solution
- stop with 6% acetic acid
- store gel in H2O
Buffers:
thiosulfate developing solution:[for 100 ml: 6 g Na2CO3, 50 µl Formaldehyde, 2 ml thiosulfate solution]
