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Protein Quantitation.....

Protein Quantitation..... - Biology Forum

Protein Quantitation..... - Biology Forums. Ask questions and discuss the study of Biology. If you have Biology questions from your homework ask them here!


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Old 01-21-2012, 08:43 AM
Pipette Filler
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Default Protein Quantitation.....



Hi, I'm working on observing a protein on a western blot, I have been unsuccessful. I have tried concentrating my sample using acetone precipitation. I'm going to try and concentrate my sample more. any suggestion?

Also, I will be starting new cell cultures using complete media consisting of DMEM w/10% FBS. Is there anyway to help my cells produce more protein?
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Old 02-06-2012, 04:20 AM
Pipette Filler
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Default Re: Protein Quantitation.....

I guess if you can't even detect your protein via Western Blot, it must be in very very low concentration or it is even not there. When I grew cells on DMEM I measured the general concentration proteins in the cell lysate just per spectrophotometry at 600 nm (standardized by using BSA standard concentrations). For IP I used about 650 g, for Western I always used about 30g of the cell material and it worked perfectly fine.
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