An improved method of sample preparation on AnchorChip targets for MALDI-MS and MS/MS and its application in the liver proteome project.
An improved method for sample preparation for MALDI-MS and MS/MS using AnchorChip targets is presented. The method, termed the SMW method (sample, matrix wash), results in better sensitivity for peptide mass fingerprinting as well as for sequencing by MS/MS than previously published methods. The method allows up-concentration and desalting directly on the mass spectrometric target and should be amenable for automation. A draw back caused by extensive oxidation of methionine and tryptophan in the SMW method can be alleviated by the addition of n-octyl glucopyranoside and DTT to the sample solution. The method was validated for protein identification from a 2-DE based liver proteome study. The SMW method resulted in identification of many more proteins and in most cases with a better score than the previously published methods.
Zhang
Xumin
X
Beijing Genomics institute, Chinese Academy of Science, Beijing, China.
Shi
Liang
L
Shu
Shaokung
S
Wang
Yuan
Y
Zhao
Kang
K
Xu
Ningzhi
N
Liu
Siqi
S
Roepstorff
Peter
P
D000818:Animals; D003460:Crystallization; D015180:Electrophoresis, Gel, Two-Dimensional; D008099:Liver / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D051379:Mice; D046208:Microchip Analytical Procedures / Q000379:methods*; D010084:Oxidation-Reduction; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D020543:Proteome