Native SDS PAGE Kit

Tool Details

*FOR RESEARCH USE ONLY

• Tool name: Native SDS PAGE Kit
• Research fields: Cell biology;Other
• Molecular formula: Sample Buffer: 100 mM Tris HCl 150 mM Tris Base 0.01875% Coomassie G-250 0.00625% Phenol Red 10% Glycerol pH 8.5 Run Buffer: 50 mM MOPS 50 mM Tris Base 0.0375% SDS pH 7.7
• Description: There is a great need for a robust method that combines the electrophoretic resolution of proteins with retention of their functional behavior. The laboratory of Dr. David Petering has devised a new method of sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in which proteins can be well separated during electrophoresis as well as maintain their native 3-dimensional conformations and functional activity. This native SDS-PAGE method will allow for important new experiments to be conducted in the proteomic field and a better understanding of proteins such as isolation from the gel of native proteins for further analysis, direct in-gel assay of enzymatic activity, direct in-gel survey of binding activity of proteins with small molecules (e.g. drugs, toxic agents) and macromolecules (e.g. protein binding partners including antibodies, cognate DNA binding sites), and isolation of protein for subsequent mass spectral identification.In traditional SDS-PAGE the proteins are well separated but are denatured such that the structure and function are no longer adequately maintained for carrying out further functional assays. The current commercial alternative, known as native PAGE, enables researchers to maintain protein function and structure, but the technique leads to poor separation and smearing of protein during electrophoresis. Our easy to implement Native SDS-PAGE method has retained the enzymatic activity of a number of enzymes tested, and has also maintained protein complexes in a bound state. This new technology will allow for multiple new applications in drug discovery, proteomics, protein therapeutics, and toxicology.
• Solubility: N/A
• Additional notes: Can be performed using commercial pre-casted gels. Simple technique and requires only one buffer system. Can retain the functional activity of proteins tested. Can retain binding of protein complexes. The neutral pH of the NSDS buffer system has an advantage over the high pH Tri-glycine system for separation of the pH sensitive proteins.

Handling

• Storage conditions: RT
• Shipping conditions: Dry Ice

References

• Nowakowski et al. 2014. Metallomics. 6(5):1068-78. PMID: 24686569.
• Native SDS-PAGE: high resolution electrophoretic separation of proteins with retention of native properties including bound metal ions.