One-step HPLC purification procedure for porcine brain 90-kDa heat shock protein.
The 90-kDa heat shock protein (HSP90) was purified from porcine brain by a novel single-step purification procedure using diethylaminoethyl high-performance liquid chromatography (HPLC). About 4.8 mg of HSP90 was isolated from 25 g wet wt porcine brain tissue. The purified protein possessed a single moiety on one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining. Western blotting using monoclonal antibody prepared against human HSP90 confirmed its identity as HSP90. These results indicate that small-scale HPLC purification of HSP90 from porcine brain tissue can be readily accomplished, with high yield, using a convenient one-step purification method. The procedure described in this paper represents a significant improvement in current purification methods for the isolation of HSP90 from porcine brain.
Chu
L F
LF
Department of Comparative Medicine, Pig Research Institute, Taiwan, Republic of China.
Lee
W C
WC
Yang
P C
PC
Chu
R
R
Huang
T Y
TY
Mao
S J
SJ
D000818:Animals; D001923:Brain Chemistry; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003720:Densitometry; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D018841:HSP90 Heat-Shock Proteins / Q000737:chemistry / Q000302:isolation & purification*; D015151:Immunoblotting; D013552:Swine
D018841:HSP90 Heat-Shock Proteins