Cell Lysis Buffer - (Oct/29/2010 )
Can someone give me a recipe for a cell lysis buffer...here is what I"m trying to do:
Detect Ras in HEK293 cells by western.
Thanks!
1% Triton x-100
50 mM Tris pH7.4
150 mM NaCl
1 mM EDTA
Protease/phosphatase inhibitors
Should do the trick.
great thanks! What is the difference between using tritonx or sds as the detergent?
I think it is their degree of "harshness", whatever that means. What method are you using for protein quantitation?
triton is non-ionic and sds is ionic
1X Cell Lysis Buffer:
20 mM Tris-HCl (pH 7.5)
150 mM NaCl
1 mM Na2EDTA
1 mM EGTA
1% Triton
2.5 mM sodium pyrophosphate
1 mM b-glycerophosphate
1 mM Na3VO4
1 μg/ml leupeptin
some protease
castleinthesky on Fri Mar 2 03:07:09 2012 said:
1X Cell Lysis Buffer:
20 mM Tris-HCl (pH 7.5)
150 mM NaCl
1 mM Na2EDTA
1 mM EGTA
1% Triton
2.5 mM sodium pyrophosphate
1 mM b-glycerophosphate
1 mM Na3VO4
1 μg/ml leupeptin
some protease
Castleinthesky, it might be worth you checking when these posts were last posted in, this one is well over a year old...