Steven Finkbeiner, Departments of Neurology and Physiology, UCSF
http://gweb1.ucsf.edu/labs/finkbeiner/Protocol/protocol_list/Lysis_Buff.htm
TGEK Base
50 mM Tris
10% vol Glycerol
1 mM EDTA
100 mM KCl
(add for std lysis buffer):
PMSF
Benzamidine
Leupeptin
Aprotinin
0.5% NP40
(add for solubilization buffer)
1% NP40
0.1% Triton X100
0.1% SDS
0.5% NaDeoxycholate
Buffer A (fractionation buffer)
10 mM Tris
1.5 mM MgCl2
10 mM KCl
Buffer C (nuclear lysis buffer)
25% glycerol
1.5 mM MgCl2
0.2 M EDTA
0.42 M NaCl
10 mM Tris
RIPA
50mM Tris HCl
150 mM NaCl
1% NP40
0.5% NaDeoxycholate
0.1% SDS
Ubi Buffer 1
(base)
50 mM Tris pH 7.4
1 mM EDTA
250 mM NaCl
0.1% NP40
1% Triton X100
0.5% SDS
0.25% DOC
(add)
100 mM NaF
5 mM NaVO3
Pepstatin
PMSF
Aprotinin
Benzamidine
Leupeptin
Ubi Buffer 2
(base)
50 mM Tris pH 7.4
1 mM EDTA
0.1% NP40
250 mM NaCl
(add)
5 mM NaF
0.5 mM NaVO3
Pepstatin
PMSF
Aprotinin
Benzamidine
Leupeptin
NP40
20 mM Tris pH 7.5
10 mM B-Glycerophosphate
2 mM EGTA
5 mM NaF
150 mM NaCl
1 mM NaPPi
1% NP40
(add)
2 mM DTT
1 mM Vanadate
.5% Aprotinin
100 uM PMSF
(or for IP, add)
Aprotinin
Pepstatin
Benzamidine
Leupeptin
PMSF
5mM Vanadate
5mM DTT
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