Hello, I want to isolate the outer membrane protein from a bacterium that I grew under different conditions and display it on sds gel. Sonication device was used in all the articles i found as a result of my research. However, there is no sonication device around me that I can reach. What can I use instead of sonication?
We have made good use of a French press for breaking cells for the purpose of isolating Gram-negative bacterial membranes. After collecting the membranes by centrifugation of the extract, the outer membranes can be prepared by dissolving the inner membranes with a detergent (N-lauroylsarcosine, I think) and centrifuging again to obtain the outer membranes. These can then be used for further analysis.
Anything that disrupts the membrane will release the desired protein. Adam B Shapiro suggests a clever method with a French press - this essentially uses high shear flow to disrupt the membrane. There are other ways to generate shear - high speed stirring for example. There are also chemical methods
to disrupt the membrane. Ultrasonic disruption is common but not the only way
French press instrument:
https://www.glenmills.com/product/french-press-g-m-high-pressure-cell-disruption/#:~:text=A%20French%20press%20is%20commonly,many%20in%20vitro%20biochemical%20assays.
Berfin Eroğlu You can try osmotic shock if your protein is outer membrane protein. Here is the link to paper that you can follow.
DOI: 10.1002/btpr.2999
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