A good and cheap alternative to SYPRO Rubi is RuBPS staining. Here are some protocols. More information can be found on www.ruthenium.ag.vu
RuBPS staining protocol I (quality)
1. Fix the gel in 30% EtOH, 10% acetic acid overnight
2. Rinse the gel in 20% EtOH for 30 min and repeat 3 times
3. Incubate the gel in 1 mM RuBPS solution for 6 h
4. Equilibrate the gel in water for 10 min and repeat once
5. Destain the gel with 40% EtOH/10% acetic acid for 15 h
6. Equilibrate the gel in water for 10 min repeat once and scan
all% are in V/V
Procedure as published in Proteomics 2004, 4, 599–608.
RuBPS staining protocol II (fast)
1. Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid containing
1 mM RuBPS for 1 h.
2. Destain the gel for 20 min in 40% Ethanol/10% acetic acid
3. Wash the gel for 10 min in water and scan
all% are in V/V
Procedure as published in PLoSONE. 2007 Feb 28;2(2)e263.
RuBPS staining protocol III (co-electrophoretical)
1. Add 1 ml of 20 mM stock solution to one pocket of your SDS gel. Run
the gel according to your standard procedure.
2. Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid for 20 min
3. Incubate the gel in 50 ml water for 10 min and scan
RuBPS staining protocol IV (loading buffer)
1. Add 1 ml of 20 mM stock solution to your loading buffer (omit all other
dyes like bromophenol blue). Run the gel according to your standard
procedure.
2. Incubate the gel in 50 ml of 40% Ethanol/10% acetic acid for 20 min
3. Incubate the gel in 50 ml water for 10 min and scan