How to Run Native Protein Gels

Biochemistry, Molecular Biology, and Cell Biology Protocols >> How to Run Native Protein Gels

1. Perform polyacrylamide gel electrophoresis as described, except do not add SDS to the running buffer or sample buffer. In addition, it may not be necessary to use a stacking gel - just the lower, resolving gel may be sufficient.

2. Bands will likely appear more smeary than normal SDS-PAGE. Therefore, run the gel at half the voltage as normal in order to have as high resolution bands as possible. Since it will probably take longer to run, try to keep the gel as cool as possible by cooling the running buffer before use.

3. For molecular weight standards, it may be a good idea to purchase a set of nondenaturing moelcular weight standards for nondenaturing gels.

4. Transfer the gel and perform blotting as described.

5. Keep in mind that in native polyacrylamide gel electrophoresis, the rate of mobility of the protein depends on both its size and its shape. Fibrillar proteins will likely run more slowly than globular proteins.