Rosebusch Tenside Buffer
12.5 mM Hepes pH 7.4
1 mM DTT
0.5 mM EGTA
100 mM KOAc
1 mM MgCl2
1% n-octylpolyoxyethylene
TNE
10 mM Tris-HCl pH 7.4
150 mM NaCl
1 mM EDTA
1% Nonidet-P40
TNTE
10 mM Tris-Hcl pH 7.5
150 mM NaCl
0.3% Triton-X100
5 mM EDTA
RIPA-buffer
10 mM Tris-Hcl pH 7.5
150 mM NaCl
1% (w/v) Triton-X 100
0.1% (w/v) SDS
1% (w/v) sodium deoxycholate
HA- Lysisbuffer (100 ml)
0.5% NP40 0.5 g
25 mM Tris pH 7.5 5 mls 0.5 M
100 mM NaCl 2 mls 5 M
add just before use 1/250 mammalian protease inhibitors
Only for Y-Phasphorylation experments:
Add 50 mM NaF (0.21 g) and 1/100 Phosphatase Inhibitor cocktail II
YP-IP Washbuffer (100 ml)
0.1 % NP40 0.1 g
25 mM Tris pH 7.5 5 mls 0.5 M
150 mM NaCl 3 mls 5 M
Only for Y-Phasphorylation experments:
add 1/100 Phosphatase Inhibitor cocktail II just before use