An on-target desalting and concentration sample preparation protocol for MALDI-MS and MS/MS analysis.
2DE coupled with MALDI-MS is one of the most widely used and powerful analytic technologies in proteomics study. The MALDI sample preparation method has been developed and optimized towards the combination of simplicity, sample-cleaning, and sample concentration since its introduction. Here we present a protocol of the so-called Sample loading, Matrix loading, and on-target Wash (SMW) method which fulfills the three criteria by taking advantage of the AnchorChip™ targets. Our method is extremely simple and no pre-desalting or concentration is needed when dealing with samples prepared from 2DE. The protocol is amendable for automation and would pave the road for high-throughput MALDI-MS or MS/MS-based proteomics studies with guaranteed sensitivity and high identification rate. The method has been successfully applied to mouse liver proteome study and so far has been employed in other proteome studies by world-wide researchers.
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Xumin
X
Zhang
Quanhui
Q
Wang
Xiaomin
X
Lou
Haidan
H
Sun
Peter
P
Roepstorff
Siqi
S
Liu
D000818:Animals; D002021:Buffers; D002138:Calibration; D003460:Crystallization; D008099:Liver / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification*; D040901:Proteomics; D012015:Reference Standards; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000592:standards; D014357:Trypsin / Q000737:chemistry
D002021:Buffers; D020543:Proteome; D014357:Trypsin