Direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometric identification of proteins on membrane detected by Western blotting and lectin blotting.
We have developed new procedures to identify proteins after they are detected by Western blotting or other interactions such as lectin blotting on membranes. Our method is based on the combination of on-membrane MALDI-TOF mass spectrometry with piezoelectric chemical inkjet technology. Using this method the GroEL, FtsZ, DnaK, and GroES proteins were successfully identified from Escherichia coli after separation on two-dimensional gels, immunostaining, and on-membrane digestion. A glycoprotein detected by lectin blotting with concanavalin A was also identified using this technique.
Ohtsu
Iwao
I
Life Science Laboratory, Analytical & Measuring Instruments Division, Shimadzu Corporation, 1 Nishinokyo-Kuwabaracho, Nakagyo-ku Kyoto 604-8511, Japan.
Nakanisi
Tsuyoshi
T
Furuta
Masaru
M
Ando
Eiji
E
Nishimura
Osamu
O
D000818:Animals; D015153:Blotting, Western / Q000295:instrumentation / Q000379:methods; D018835:Chaperonin 10 / Q000032:analysis; D018834:Chaperonin 60 / Q000032:analysis; D003208:Concanavalin A / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli; D029968:Escherichia coli Proteins / Q000032:analysis*; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis; D037102:Lectins / Q000032:analysis*; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D018835:Chaperonin 10; D018834:Chaperonin 60; D029968:Escherichia coli Proteins; C076835:FtsZ84 protein, E coli; D018840:HSP70 Heat-Shock Proteins; D037102:Lectins; D011506:Proteins; D003208:Concanavalin A; C063842:dnaK protein, E coli