Abstract
The efficient extraction of proteins of interest from cells and tissues can be challenging. Here we demonstrate the differences in extraction of the focal adhesion protein Kindlin-2 and the transcriptional repressor Snail from choriocarcinoma cells using NP-40 and RIPA lysis buffer. We also show the use of a more denaturing urea/thiourea lysis buffer for solubilization, by comparing its effectiveness with the often utilized RIPA lysis buffer for solubilization of heat shock proteins (HSP) B1 and B5 and the cytoplasmic adapter protein integrin-linked kinase (ILK) from smooth muscle. Overall, the results demonstrate the importance of optimizing lysis buffers for specific protein solubilization prior to finalizing the experimental workflow.
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Acknowledgment
This work was supported by a Natural Sciences and Engineering Research Council Discovery Grant (#250218), an Establishment Grant from the Saskatchewan Health Research Foundation (SHRF; #2695), and a regional partnership program grant from SHRF (#2776) and the Canadian Institutes of Health Research (#ROP-101051) to DJM.
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Miskiewicz, E.I., MacPhee, D.J. (2019). Lysis Buffer Choices Are Key Considerations to Ensure Effective Sample Solubilization for Protein Electrophoresis. In: Kurien, B., Scofield, R. (eds) Electrophoretic Separation of Proteins. Methods in Molecular Biology, vol 1855. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8793-1_5
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DOI: https://doi.org/10.1007/978-1-4939-8793-1_5
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