SDS sample buffer

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==Protein gel sample loading buffer==
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==Purpose==
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===Purpose===
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* For preparation and loading of protein samples onto a gel for SDS-PAGE analysis.
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*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
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===Use===
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==Use of loading buffer==
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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.
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===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]
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===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 09:40, 27 October 2011

Contents

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

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