protein extraction from primary cells - (Jul/07/2009 )
Hi all,
Im working with HUAECs, primary cells. Some of the proteins I'm interested in can be found in the cell and others are released into the medium during the growth of the cells in culture.
Could anyone advise me which methods are best to use to extract both types of proteins from these cells. The process needs to be compatible with the BCA assay and Western Blotting.
I have used Pierce M-Per reagent for lysing the proteins from the cells, I saved the medium from the flask but I don't know what method would be best or if even the M-Per one is good enough for the cells.
Many thanks.
-kellyly-
kellyly on Jul 7 2009, 05:43 AM said:
Hi all,
Im working with HUAECs, primary cells. Some of the proteins I'm interested in can be found in the cell and others are released into the medium during the growth of the cells in culture.
Could anyone advise me which methods are best to use to extract both types of proteins from these cells. The process needs to be compatible with the BCA assay and Western Blotting.
I have used Pierce M-Per reagent for lysing the proteins from the cells, I saved the medium from the flask but I don't know what method would be best or if even the M-Per one is good enough for the cells.
Many thanks.
Im working with HUAECs, primary cells. Some of the proteins I'm interested in can be found in the cell and others are released into the medium during the growth of the cells in culture.
Could anyone advise me which methods are best to use to extract both types of proteins from these cells. The process needs to be compatible with the BCA assay and Western Blotting.
I have used Pierce M-Per reagent for lysing the proteins from the cells, I saved the medium from the flask but I don't know what method would be best or if even the M-Per one is good enough for the cells.
Many thanks.
For protein isolation from culture media see the first protocol. For isolation of proteins from primary cells, it is as good as any of the many protein extraction protocols. I use the NP40 Lysis method, but M-Per, RIPA all are good.
-cellcounter-