Utility of dry gel from two-dimensional electrophoresis for peptide mass fingerprinting analysis of silkworm proteins.
We compared the use of wet and dry two-dimensional electrophoresis (2-DE) gels for in-gel tryptic digestion and subsequent analysis by mass spectrometry, first using bovine serum albumin (BSA) as a model protein and then using unknown proteins from an extract of the silkworm midgut. The gel was either dried at 80 degrees C or left wet. Upon analysis of BSA, there was little difference in peptide recovery from 2-DE or in mass spectrum between the dry and the wet gels. The midgut extract was resolved into more than 1,100 protein spots by 2-DE, and 40 of these spots were sampled for further analysis. For all of the 40 proteins, the results obtained from dry and wet gels were quite similar in mass spectra and protein identification, although the relative amounts of peptides from tryptic digestion ranged from 45 to 146%. Based on these results, we confirmed the utility of dry electrophoretic gels for proteomics of insect extracts.
Zhang
Pingbo
P
Laboratory of Insect Genetic Resources, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan.
Yamamoto
Kohji
K
Wang
Yongqiang
Y
Banno
Yutaka
Y
Fujii
Hiroshi
H
Miake
Fumio
F
Kashige
Nobuhiro
N
Aso
Yoichi
Y
D000818:Animals; D012831:Bombyx / Q000378:metabolism*; D002417:Cattle; D015180:Electrophoresis, Gel, Two-Dimensional; D005782:Gels / Q000737:chemistry; D019476:Insect Proteins / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D020543:Proteome; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D005782:Gels; D019476:Insect Proteins; D010455:Peptides; D020543:Proteome; D012710:Serum Albumin, Bovine