Coloração Proteínas em Gel

WHICH REACTION EQUATION OF CONVERT Cu+2 TO Cu+1 in the protein test

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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

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Thank yoy