SDS sample buffer

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
m (Changed name of categories from plural to singular.)
(SDS link)
Line 5: Line 5:
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  
-
*0.8 g SDS  
+
*0.8 g [[SDS]]
*4.0 ml 100% glycerol  
*4.0 ml 100% glycerol  
*0.4 ml 14.7 M β-mercaptoethanol  
*0.4 ml 14.7 M β-mercaptoethanol  
Line 22: Line 22:
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 10:51, 8 April 2009

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

Personal tools