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problem in Protein qunatification : mice carotid artery

problem in Protein qunatification : mice carotid artery - Biology Forum

problem in Protein qunatification : mice carotid artery - 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 08-29-2011, 07:37 PM
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Default problem in Protein qunatification : mice carotid artery



Hai,

I am doing the rsearch in cardiovascular system. In my project, I am trying to qunatify the protein content from mice carotid artery. My problem is, After quanitfication, when I ran the gel, I didnt get the equal band size for all the sample ( with commasiev staining as well as in the Western blot)...

I have used the 2x SDS buffer for lysis the tissue and then centrifuged @ 14000 rpm for 15 min. After that I took the supernatant for the protein qunatification. I did the protein quantification with BCA protein assay then based on the value I have loaded 10 microgram (Protein) of sample to each well.

I have tried various steps and repeated 3-4 times ... I feel there is a problem in lysis buffer...!!!! I want to know the buffer which I am using (2 x SDS) is correct or not.....!!! I need some idea for you to solve this problem.

with Regards,

Saran
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Old 08-31-2011, 05:23 PM
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Default Re: problem in Protein qunatification : mice carotid artery

High SDS interferes with the BCA protein assay. Manuf. instructions say 5% SDS or less is compatible. So check it and dilute if necessary.
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