Dear Scientists,
I am trying to check some protein expression in yeast using western blot technique, but I got stuck for protein extraction from Yeast. What is the difference between the lysis for cell and yeast?
You can please follow the links below:-
1. http://onlinelibrary.wiley.com/doi/10.1002/yea.1905/pdf
2. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2031916/
Yeast is very difficult to be broken. You may try freeze thawing for several times before protein extraction.
Dear Lalita
There is no differences between cells lysis of yeast or bacteria if I got you right! The whole idea is to get internal contents that include protein, for yeast the easiest (to me at least) method is freezing & grinding in liquid nitrogen (freeze cell in liquid nitrogen then ground frozen cell into powder using a pestle & mortar that chilled in liquid nitrogen as well, the good thing about this method that you can keep this powder at -80 C degree, for indefinite, for cell lysate add five volume of buffer to this powder. You can read all the methods used in yeast cell lysis the following link.
https://www.embl.de/pepcore/pepcore_services/protein_purification/extraction_clarification/cell_lysates_yeast/
Hope this helps
The main question is : do you want to harvest active protein or to have an idea about the amount of proteins? In the first case you can use the grinding with glass beads after freezing in liquid nirtogen , or also an other possibility is the French press... In the past some enzymes were used to digest more or less the cell wall before the grinding in order to increase the % of cell lysis, however a decrese in some yeast enzymic activities of the extracts could be more or less lost or decreased. Anyways, good luck
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