SDS sample buffer: Difference between revisions
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==Purpose== | ==Purpose== | ||
* For preparation and loading of protein samples onto a gel for SDS-PAGE analysis. | * For preparation and loading of protein samples onto a gel for SDS-PAGE analysis. | ||
==Preparation== | ==Preparation== | ||
To make 10 mL of | ====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]] | ||
| Line 11: | Line 12: | ||
*8 mg bromophenol Blue | *8 mg bromophenol Blue | ||
Final concentrations ( | ====Final concentrations (1x)==== | ||
*50 mM Tris-HCl pH 6.8 | *50 mM Tris-HCl pH 6.8 | ||
*2% [[SDS]] | *2% [[SDS]] | ||
| Line 18: | Line 19: | ||
*12.5 mM EDTA | *12.5 mM EDTA | ||
*0.02 % bromophenol blue | *0.02 % bromophenol blue | ||
==Use of loading buffer== | ==Use of loading buffer== | ||
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 06:41, 27 October 2011
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 of loading buffer
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.
