Hello, today i want to ask about ripa buffer, buffer used in
Western blot.(Very simple but seemingly important question)
You may already know this, but i found out that
there are numerous recipes for making ripa buffer.
So i concluded that researchers follow recipes of ripa buffer
which suit their experiments the best and there is no absolute
recipe.
So similarly, as for my research, should i evaluate whether
specific recipe suits my work or not based on final outcomes
of western blot because no standardized recipe exists?
Thank you for your interest and good luck with your research!
Dear Lee!
Here is the RIPA recipe. Hopefully is useful for you!
RIPA buffer (05-01)
10 mM Tris-Cl (pH 8.0)
1 mM EDTA
0.5 mM EGTA
1% Triton X-100
0.1% sodium deoxycholate
0.1% SDS
140 mM NaCl
The above solution is stable at room temperature.
Before use, add:
1 mM PMSF
Dear Lee!
Here is the RIPA recipe. Hopefully is useful for you!
RIPA buffer (05-01)
10 mM Tris-Cl (pH 8.0)
1 mM EDTA
0.5 mM EGTA
1% Triton X-100
0.1% sodium deoxycholate
0.1% SDS
140 mM NaCl
The above solution is stable at room temperature.
Before use, add:
1 mM PMSF
Hi Lee,
The RIPA buffer recipe that is working well for me at the moment:
140 mM NaCl
25 mM Tris-HCl
1 mM EDTA
0.5 mM EGTA
1 mM PMSF
1mM NaF
1% Triton X-100
0.1% SDS
0.1% Protease inhibitors cocktail
Ultrapure water. PH adjusted to 7.4.
Regards.
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