SDS sample buffer

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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.
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==Preparation==
==Preparation==
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To make 10 mL of 4X stock:
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====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]]  
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*8 mg bromophenol Blue  
*8 mg bromophenol Blue  
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Final concentrations (1X):
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====Final concentrations (1x)====
*50 mM Tris-HCl pH 6.8  
*50 mM Tris-HCl pH 6.8  
*2% [[SDS]]
*2% [[SDS]]
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*12.5 mM EDTA  
*12.5 mM EDTA  
*0.02 % bromophenol blue
*0.02 % bromophenol blue
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==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.
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==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]
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==References==
==References==

Revision as of 09:41, 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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