2 replies to this topic

[Alfred Nobel]

I want to extract protein from small numbers of cancer cells using RIPA buffer plus sonication. My question concerns how much power (watts) is needed to adequately disrupt the DNA so the slime does not interfere with my protein assay and subsequent gel electrophoresis. I am using volumes of around 50-100 ul/extraction.
Thanks

[cellcounter]

Alfred Nobel, on Aug 6 2009, 10:17 AM, said:

I want to extract protein from small numbers of cancer cells using RIPA buffer plus sonication. My question concerns how much power (watts) is needed to adequately disrupt the DNA so the slime does not interfere with my protein assay and subsequent gel electrophoresis. I am using volumes of around 50-100 ul/extraction.
Thanks

Sonication power needed is a function of what sonicator you are using, what number of cells, and what cells.

So, the best thing to do is to optimize the conditions for yourself using the same small number of cells, in the same volume, at different power and different time intervals on your machine. Do the protein assay, and whichever worked the best, use that for real tests.

Edited by cellcounter, 06 August 2009 - 11:01 AM.

[Alfred Nobel]

cellcounter, on Aug 6 2009, 12:01 PM, said:

Alfred Nobel, on Aug 6 2009, 10:17 AM, said:

I want to extract protein from small numbers of cancer cells using RIPA buffer plus sonication. My question concerns how much power (watts) is needed to adequately disrupt the DNA so the slime does not interfere with my protein assay and subsequent gel electrophoresis. I am using volumes of around 50-100 ul/extraction.
Thanks

Sonication power needed is a function of what sonicator you are using, what number of cells, and what cells.

So, the best thing to do is to optimize the conditions for yourself using the same small number of cells, in the same volume, at different power and different time intervals on your machine. Do the protein assay, and whichever worked the best, use that for real tests.

Well I failed to mention that I am shopping for a sonicator and don't want to buy one that's too weak. The one I can afford is 150 watts max.