Improving sodium dodecyl sulfate polyacrylamide gel electrophoresis detection of low-abundance protein samples by rapid freeze centrifugation.
This work presents a rapid and simple freeze centrifugation method to concentrate dilute protein solutions for detection by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) Coomassie blue staining. Moreover, a simple way to assemble a cryoconcentration device is presented, and its use is discussed. Commercial purified protein standard and an enzyme with high fructosyltransferase (FTase) activity, coming from target fractions obtained by chromatographic separation, were used as an example. FTase, coming directly from the chromatographic fractions, was difficult to view through SDS-PAGE analysis; however, it was easily visualized, and its activity was enhanced, after the application of the freeze centrifugation protocol presented here.
Bioengineering Laboratory, University of Colima, Coquimatlán, Col 28400, Mexico; Faculty of Chemical Science, University of Colima, Coquimatlán, Col 28400, Mexico.
J J
JJ
Virgen-Ortíz
V
V
Ibarra-Junquera
P
P
Escalante-Minakata
J A
JA
Osuna-Castro
J de J
Jde J
Ornelas-Paz
N A
NA
Mancilla-Margalli
R L
RL
Castañeda-Aguilar
D000818:Animals; D001230:Aspergillus / Q000201:enzymology; D002498:Centrifugation / Q000295:instrumentation*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004867:Equipment Design; D005615:Freezing; D006602:Hexosyltransferases / Q000302:isolation & purification; D006736:Horses; D011506:Proteins / Q000302:isolation & purification*; D001426:Bacterial Proteins
D006602:Hexosyltransferases; D011506:Proteins; C021451:inulosucrase; D001426:Bacterial Proteins
Chromatographic fraction; Concentrating of proteins; Cryoconcentrator device; Freeze centrifugation; SDS–PAGE