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Laemmli-SDS-PAGE
Published: Jun 5, 2011 DOI: 10.21769/BioProtoc.80 Views: 161265
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Abstract
Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is used to separate proteins with relative molecular mass no smaller than 10 KD. Very small proteins (<10 KD) are difficult to resolve due to low ability of binding to SDS, which can be solved by gradient gels or using different eletrophoresis conditions, like Tricine-SDS-page. The basic Laemmli SDS PAGE procedure is described here.
Keywords: SDS-PAGE
Protein
Electrophoresis
Materials and Reagents
- Pre-stain Protein MW marker (Bio-Rad Laboratories)
- TEMED (Life Technologies, Gibco®)
- Ammonium persulfate (Sigma-Aldrich)
- SDS (Research Organics INC)
- 30% Acrylamide stock (37.5: 1 acrylamide: bisacrylamide) (Bio-Rad Laboratories)
- Bromophenol Blue (Thermo Fisher Scientific)
- Tris Base (Calbiochem-Behring)
- Glycine (EM Science)
- EDTA (Amresco)
- Glycerol (EM Science)
- Isopropanol
- Tris-HCl (pH 6.8)
- β-mercaptoethanol (Sigma-Aldrich)
- 10x running buffer (see Recipes)
- 2x SDS protein sample buffer (see Recipes)
Equipment
- Protein mini gel cassettes (Bio-Rad)
- Heating block module
- Table-top centrifuge
- Power supply
- Gloves
- Filter paper
Procedure
Category
Biochemistry > Protein > Electrophoresis
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