A novel droplet-tap sample-loading method for two-dimensional gel electrophoresis.
A novel procedure, droplet-tap mode, has been devised for sample application for two-dimensional gel electrophoresis (2DE) expression profiles. The sample was loaded by evenly distributed tapping of droplets of the sample on to the rehydration buffer (RB) and then lowering the strip on to the solution surface. At normal loading concentrations, the number of spots obtained was increased by approximately one-third by this new approach compared with the rehydration loading procedure. The method also resulted in significantly improved resolution compared with cup loading when high concentrations of proteins were present, indicating its potential usefulness in micropreparative separation. In addition, recovery of the proteins confirmed that protein uptake was enhanced by use of this method. By enabling improved performances in 2DE, the proposed procedure has much potential for sample loading to meet the requirements of global proteome analysis.
Long
X-H
XH
College of Life Science, Zhejiang University, Hangzhou, 310029, P.R. China.
Lin
J-F
JF
Zhang
Y-Z
YZ
D000818:Animals; D012831:Bombyx; D002623:Chemistry Techniques, Analytical / Q000295:instrumentation* / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation* / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D019476:Insect Proteins / Q000737:chemistry; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D011506:Proteins / Q000032:analysis*; D020543:Proteome; D040901:Proteomics / Q000379:methods
D019476:Insect Proteins; D011506:Proteins; D020543:Proteome