Abstract

 

DIRECT MATRIX-ASSISTED LASER DESORPTION/IONIZATION TIME-OF-FLIGHT MASS SPECTROMETRIC IDENTIFICATION OF PROTEINS ON MEMBRANE DETECTED BY WESTERN BLOTTING AND LECTIN BLOTTING

Iwao Ohtsu, Tsuyoshi Nakanisi, Masaru Furuta, Eiji Ando, Osamu Nishimura 



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.



Reproduced with permission from Journal of Proteome Research (2005)

vol. 4, pp. 1391-1396. Copyright 2005 Am.Chem.Soc.

(Published on Web: June 21, 2005)

 

 

DIRECT MS/MS ANALYSIS OF PROTEINS BLOTTED ON MEMBRANES BY A MATRIX-ASSISTED LASER DESORPTION/IONIZATION-QUADRUPOLE ION TRAP-TIME-OF-FLIGHT TANDEM MASS SPECTROMETER

Tsuyoshi Nakanishi, Iwao Ohtsu, Masaru Furuta, Eiji Ando, and Osamu Nishimura 



We constructed a system for the microscale identification of membrane-blotted proteins by proteolytic digestion using an instrument developed with piezoelectric chemical inkjet technology and MS/MS analyses of the resulting peptides with a matrix-assisted laser desorption/ionization-quadrupole ion trap-time-of-flight tandem mass spectrometer (MALDI-QIT-TOF MS). Using this system, bovine serum albumin was clearly identified at levels less than 100 fmol, and proteins from an Escherichia coli extract were also identified by an MS/MS ion search.



Reproduced with permission from Journal of Proteome Research (2005)

vol. 4, pp. 743-747. Copyright 2005 Am.Chem.Soc.

(Published on Web: April 05, 2005)

 

 

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