Activity 3: Guide
Guide Questions and References
Guide Questions
- Why is there a need for a
different resolving and stacking gel?
- How does Coomassie staining work? Are there other alternatives to this?
- Construct a standard curve using the protein profile of your protein standards. Can you estimate the weight of distinct bands from the gel?
- Give other methods that will enable visualization of isolated proteins.
References
Corpas, F. J., Carreras, A., Esteban, F. J., Chaki, M., Valderrama, R., Río, L. A., & Barroso, J. B. (2008). Localization of S‐Nitrosothiols and Assay of Nitric Oxide Synthase and S‐Nitrosoglutathione Reductase Activity in Plants. Globins and Other Nitric Oxide-Reactive Proteins, Part B Methods in Enzymology, 561-574. doi:10.1016/s0076-6879(07)37028-6
Scott, V., Clark, A. R., & Docherty, K. (n.d.). The Gel Retardation Assay. Protocols for Gene Analysis, 339-348. doi:10.1385/0-89603-258-2:339
Walker, J. M. (2009). Nondenaturing Polyacrylamide Gel Electrophoresis of Proteins. Springer Protocols Handbooks The Protein Protocols Handbook, 171-176. doi:10.1007/978-1-59745-198-7_20
Walker, J. M. (2009). SDS Polyacrylamide Gel Electrophoresis of Proteins. Springer Protocols Handbooks The Protein Protocols Handbook, 177-185. doi:10.1007/978-1-59745-198-7_21
Wilhelm, E., Takacs, C., & Bell, B. (2011). Probing Endogenous RNA Polymerase II Pre-initiation Complexes by Electrophoretic Mobility Shift Assay. Methods in Molecular Biology Transcriptional Regulation, 63-74. doi:10.1007/978-1-61779-376-9_4