RIPA buffer mistake - (Jul/06/2006 )
I made my RIPA buffer with to a final concentration of 1% Tween instead of 1% triton-x by mistake. I used this solution to extract protein from a time course experiment. Is there any point trying to run it on a gel to see if I get anything, or is all hope lost?
-Paramyxo-
QUOTE (Paramyxo @ Jul 6 2006, 04:33 PM)
I made my RIPA buffer with to a final concentration of 1% Tween instead of 1% triton-x by mistake. I used this solution to extract protein from a time course experiment. Is there any point trying to run it on a gel to see if I get anything, or is all hope lost?
Triton is basically a detergent that helps in getting the protein out of the cells. Your protein extraction is not going to that good. However, the SDS that is usually in RIPA buffer may get some of ypour protein out.
If I were you....i would throw it out. Saves time and money
-Casper-
As u might do the rest of ur experiments with 1% Tween, u might as well repeat the present experiment with new buffer (containing Tween).
If u would run it & try WB, U will see something depending if the protein will b extracted by triton.
-scolix-