I am interested in protein isolation from Saccharomyces cerevisiae. But I do not know detailed list of protein of S.cerevisiae. Should I focus on protein extract isolation or single protein isolation ? How should I use western blotting technique to isolate yeast protein(s) ? Expert suggestions are expected. Thanks.
Can you rephrase the question? It is not clear what is it exactly that you need.
If you want a list of yeast proteins head over to uniprot.org and search their proteome collection. You can start here: http://www.uniprot.org/proteomes/UP000002311
You cannot use Western blotting to isolate any protein. It is not a purification technique.
Whether you focus on preparing protein extracts or purifying single proteins depends on what you want.
Hope it helps!
Thanks Alejandro Martin for your reply.
Let me rephrase the questions. First, I wanted to know what are proteins of S.cerevisiae. Then I asked about protein isolation from S.cerevisiae in which how could I isolate single protein/whole protein.
And my bad, I asked WBT about isolation, it would have been about "identification". I never done WBT before, does this method uses primers like used in PCR ? Any hints about universal primers ! Thanks again for helping me out.
Hi Amin, Western Blots are used with antibodies to detect proteins for example in a extract. To purify a specific protein you can produce a recombinant one in bacteria to put some N or C-terminal tag. Then, you can purify using some chromatography technique. Primers are useful just to work with DNA and not proteins. So, if you want to work with proteins you will need antibodies.
To isolate a specific protein is not so easy, you'll have to work a lot. I reccomend you to read some textbooks to learn more about molecular biology techniques.
Hope it helps!
Raphael
Thank you Raphael Pavani. I totally forgot the basics. By the way that was very helpful.
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