Cytoskeletal bound proteins extract buffer:
10 mM Tris, pH 7.4
100 mM NaCl
1 mM EDTA
1 mM EGTA
1 mM NaF
20 mM Na₄P₂O₇
2 mM Na₃VO₄
1% Triton X-100
10% glycerol
0.1% SDS
0.5% deoxycholate

Soluble protein buffer:
20 mM Tris-HCl, pH 7.5
1 mM EGTA (Ca²⁺ chelator)

RIPA buffer (radio immuno precipitation assay) buffer:
RIPA buffer contains the ionic detergent sodium deoxycholate as an active constituent and is particularly used for nuclear membrane disruption for nuclear extracts. A RIPA buffer gives low background but can denature kinases. It can also disrupt protein-protein interactions (and may therefore be problematic for immunoprecipitations/pull down assays).

50 mM Tris HCl pH 8
150 mM NaCl
1% NP-40
0.5% sodium deoxycholate
0.1% SDS

The 10% sodium deoxycholate stock solution (5 g into 50 ml) must be protected from light.

The 100 mM EDTA stock solution is made with 1.86 g into 40 ml H2O and then add NaOH to dissolve and adjust pH to 7.4. Finally, adjust the total volume to 50 ml). Store the buffer at 4°C.

Nonidet-P40 (NP-40) buffer:
20 mM Tris HCl pH 8
137 mM NaCl
10% glycerol
1% nonidet P-40
2 mM EDTA

Sodium orthovanadate preparation:
This needs to be done under the fume hood

1. Prepare a 100 mM solution in double distilled water
2. Set pH to 9.0 with HCl
3. Boil until colorless
4. Cool to room temperature
5. Set pH to 9.0 again
6. Boil again until colorless
7. Repeat this cycle until the solution remains at pH 9.0 after boiling and cooling
8. Bring up to the initial volume with water
9. Store in aliquots at -20°C