Protein Isolation Workflow
Description
Scheme of Protein Isolation Workflow
1. Lyse using RIPA buffer
2. Precipitation of protein pellet
3. Lipid removal
4. Methanol wash and air-dry overnight
5. 0.1% RapiGest detergent
6. Dithiothreitol at 60°C
7. 2-iodoacetamide at RT
8. Trypsin for 16 hours at room temperature
9. 0.4% aqueous trifluoroacetic acid precipitation
10. Total recovery vials
Acknowledgements
References
Joglekar, M. V., & Hardikar, A. A. (2012). Isolation, expansion, and characterization of human islet-derived progenitor cells. Methods in molecular biology (Clifton, N.J.), 879, 351–366. https://doi.org/10.1007/978-1-61779-815-3_21
Wong, W. K. M., Joglekar, M. V., Saini, V., Jiang, G., Dong, C. X., Chaitarvornkit, A., Maciag, G. J., Gerace, D., Farr, R. J., Satoor, S. N., Sahu, S., Sharangdhar, T., Ahmed, A. S., Chew, Y. V., Liuwantara, D., Heng, B., Lim, C. K., Hunter, J., Januszewski, A. S., Sørensen, A. E., … Hardikar, A. A. (2021). Machine learning workflows identify a microRNA signature of insulin transcription in human tissues. iScience, 24(4), 102379. https://doi.org/10.1016/j.isci.2021.102379
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