Re-extracting proteins from pellet

Use this category for questions regarding problems manipulating proteins in molecular biology applications (expression, detection, etc.)

Moderators: mdfenko, leekaming

Re-extracting proteins from pellet

Postby newbie17425 » Feb 23 2017 8:51 am

Hi,

I am extracting proteins from rat hippocampus and using it for Western Blot (WB). I'm using RIPA buffer to extract the proteins, however, I'm out of my protein sample now. I saved the pellet, from my 1st extraction. Can I use the pellet again to re-extract proteins using the same method as I did before, i.e. adding RIPA and Halt-protease inhibitor?

Thanks.
newbie17425
Prolific Post-Master
Prolific Post-Master
 
Posts: 116
Joined: May 22 2015 3:23 pm

Re: Re-extracting proteins from pellet

Postby mdfenko » Feb 23 2017 11:03 am

if your first extraction was good then a second extraction will probably not give a good enough yield.
talent does what it can
genius does what it must
i do what i get paid to do
mdfenko
ModSquad
ModSquad
 
Posts: 1461
Joined: Jan 24 2006 2:39 pm
Location: Staten Island, NY USA

Re: Re-extracting proteins from pellet

Postby newbie17425 » Feb 23 2017 11:17 am

But in principle it is possible, assuming yields from my proteins from the 1st extraction was not so good.
newbie17425
Prolific Post-Master
Prolific Post-Master
 
Posts: 116
Joined: May 22 2015 3:23 pm

Re: Re-extracting proteins from pellet

Postby mdfenko » Feb 23 2017 2:45 pm

if there are any intact cells in the tissue then you may be able to extract again. however, if cellular disruption was efficient, then you will not be able to extract efficiently.
talent does what it can
genius does what it must
i do what i get paid to do
mdfenko
ModSquad
ModSquad
 
Posts: 1461
Joined: Jan 24 2006 2:39 pm
Location: Staten Island, NY USA


Return to Protein Methods

Who is online

Users browsing this forum: No registered users and 1 guest