SDS sample buffer: Difference between revisions
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To make 10 mL of 4X stock: | To make 10 mL of 4X stock: | ||
* 2.0 ml 1M Tris-HCl pH 6.8 | * 2.0 ml 1M Tris-HCl pH 6.8 | ||
*0.8 g SDS | *0.8 g [[SDS]] | ||
*4.0 ml 100% glycerol | *4.0 ml 100% glycerol | ||
*0.4 ml 14.7 M β-mercaptoethanol | *0.4 ml 14.7 M β-mercaptoethanol | ||
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Dilute protein sample 1:3 into 4X sample loading buffer. Load on acrylimide gel in SDS-PAGE buffer. | Dilute protein sample 1:3 into 4X sample loading buffer. Load on acrylimide gel in SDS-PAGE buffer. | ||
===Safety=== | ===Safety=== | ||
β-mercaptoethanol is a severe irritant and is readily absorbed through the skin. SDS is a respiratory irritant in solid form and a mask should be worn while weighing it. | β-mercaptoethanol is a severe irritant and is readily absorbed through the skin. [[SDS]] is a respiratory irritant in solid form and a mask should be worn while weighing it. | ||
*[http://ptcl.chem.ox.ac.uk/MSDS/ME/2-mercaptoethanol.html β-mercaptoethanol MSDS] | *[http://ptcl.chem.ox.ac.uk/MSDS/ME/2-mercaptoethanol.html β-mercaptoethanol MSDS] | ||
===References=== | ===References=== | ||
Revision as of 07:51, 8 April 2009
Protein gel sample loading buffer
Purpose
- For preparation and loading of protein samples onto a gel for SDS-PAGE analysis.
Preparation
To make 10 mL of 4X stock:
- 2.0 ml 1M Tris-HCl pH 6.8
- 0.8 g SDS
- 4.0 ml 100% glycerol
- 0.4 ml 14.7 M β-mercaptoethanol
- 1.0 ml 0.5 M EDTA
- 8 mg bromophenol Blue
Final concentrations (1X):
- 50 mM Tris-HCl pH 6.8
- 2% SDS
- 10% glycerol
- 1% β-mercaptoethanol
- 12.5 mM EDTA
- 0.02 % bromophenol blue
Use
Dilute protein sample 1:3 into 4X sample loading buffer. Load on acrylimide gel in SDS-PAGE buffer.
Safety
β-mercaptoethanol is a severe irritant and is readily absorbed through the skin. SDS is a respiratory irritant in solid form and a mask should be worn while weighing it.
