SDS sample buffer

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(Preparation)
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* 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 10% glycerol  
+
*4.0 ml 100% glycerol  
*0.4 ml 14.7 M β-mercaptoethanol  
*0.4 ml 14.7 M β-mercaptoethanol  
*1.0 ml 0.5 M EDTA  
*1.0 ml 0.5 M EDTA  
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*12.5 mM EDTA  
*12.5 mM EDTA  
*0.02 % bromophenol blue
*0.02 % bromophenol blue
 +
===Use===
===Use===
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.

Revision as of 12:56, 13 March 2008

Contents

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.

References

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