SDS sample buffer: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(SDS link) |
mNo edit summary |
||
| Line 1: | Line 1: | ||
==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. | |||
==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 | ||
| Line 19: | Line 19: | ||
*0.02 % bromophenol blue | *0.02 % bromophenol blue | ||
==Use of loading buffer== | |||
Dilute protein sample 1:3 into 4X sample 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. | 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== | |||
*[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.
