Difference between revisions of "SDS sample buffer"

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==Protein gel sample loading buffer==
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==Purpose==
===Purpose===
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* 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.
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===Preparation===
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==Preparation==
 
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  
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*0.02 % bromophenol blue
 
*0.02 % bromophenol blue
  
===Use===
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==Use of loading buffer==
Dilute protein sample 1:3 into 4X sample loading buffer. Load on acrylimide gel in SDS-PAGE buffer.
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Dilute protein sample 1:3 into 4X sample loading buffer. Load on acrylimide gel in SDS-PAGE buffer.
===Safety===
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 +
==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===
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 +
==References==
 
*[http://genetics.mgh.harvard.edu/szostakweb/resources/Public%20Protocols/protein_page/index.html Szostak Lab SDS-PAGE]
 
*[http://genetics.mgh.harvard.edu/szostakweb/resources/Public%20Protocols/protein_page/index.html Szostak Lab SDS-PAGE]
 
[[Category:Material]]
 
[[Category:Material]]
 
[[Category:Buffer]]
 
[[Category:Buffer]]

Revision as of 06:40, 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.

References