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RIPA buffer contents - comparison of 2 buffers (Jan/12/2009 )

i have recipes of 2 different RIPA buffers:

1.)
. Tris-HCl: 50 mM, pH 7.4
· NP-40: 1%
· Na-deoxycholate: 0.25%

· NaCl: 150 mM
· EDTA: 1 mM

+Protease Inhibitors

2.)
• 50 mM Tris-HCl pH 7.4
• 150 mM NaCl
• 1% Triton x-100
• 1% Sodium deoxycholate
• 0.1% SDS

• 1 mM EDTA

+Protease Inhibitors

Q: what is the differece of these 2 buffers? could the same proteins be extracted by this buffers?

-moljul-

QUOTE (moljul @ Jan 12 2009, 10:25 AM)
i have recipes of 2 different RIPA buffers:

1.)
. Tris-HCl: 50 mM, pH 7.4
· NP-40: 1%
· Na-deoxycholate: 0.25%

· NaCl: 150 mM
· EDTA: 1 mM

+Protease Inhibitors

2.)
• 50 mM Tris-HCl pH 7.4
• 150 mM NaCl
• 1% Triton x-100
• 1% Sodium deoxycholate
• 0.1% SDS

• 1 mM EDTA

+Protease Inhibitors

Q: what is the differece of these 2 buffers? could the same proteins be extracted by this buffers?

I think the first one is also called NP40 lysis buffer (or modified-ripa buffer to some), while the second with triton-x 100 is original one.

The difference is in the detergent used (no40/triton), and I believe ripa (with triton) is stronger.

-cellcounter-

QUOTE (cellcounter @ Jan 12 2009, 07:06 PM)
QUOTE (moljul @ Jan 12 2009, 10:25 AM)
i have recipes of 2 different RIPA buffers:

1.)
. Tris-HCl: 50 mM, pH 7.4
· NP-40: 1%
· Na-deoxycholate: 0.25%

· NaCl: 150 mM
· EDTA: 1 mM

+Protease Inhibitors

2.)
• 50 mM Tris-HCl pH 7.4
• 150 mM NaCl
• 1% Triton x-100
• 1% Sodium deoxycholate
• 0.1% SDS

• 1 mM EDTA

+Protease Inhibitors

Q: what is the differece of these 2 buffers? could the same proteins be extracted by this buffers?

I think the first one is also called NP40 lysis buffer (or modified-ripa buffer to some), while the second with triton-x 100 is original one.

The difference is in the detergent used (no40/triton), and I believe ripa (with triton) is stronger.


Yes I agree. The first has two detergents (nonyl-phenoxylpolyethoxylethanol and sodium deoxycholate). The second additionally SDS and Triton but no NP40. Sodium deoxycholate and Na-deoxycholate are the same detergent (sodium = natrium), only the concentrations are different.

-hobglobin-