Two-dimensional gel electrophoresis characterization of the mouse leukocyte proteome, using a tri-reagent for protein extraction.
The use of "tri-reagents" allows the concomitant isolation of DNA, RNA, and proteins for a complete molecular characterization of biological samples. The aim of the current study was to perform the comparative evaluation of two-dimensional gel proteomes isolated with or without a tri-reagent protein extraction step before dissolving the samples in the first-dimension electrophoresis buffer. We conclude that the use of tri-reagents increases the amount of protein extracted from the sample. Furthermore, an average of 301 +/- 3.6 spots were found in gels from both sample preparation methods, whereas 71.7 +/- 8.1 and 49.7 +/- 2.3 spots were uniquely seen in tri-reagent and non-tri-reagent samples, respectively.
Lenchik
Nataliya I
NI
Department of Medicine, University of Tennessee Health Science Center, Memphis, 38104, USA.
Desiderio
Dominic M
DM
Gerling
Ivan C
IC
D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D007962:Leukocytes / Q000737:chemistry*; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D008970:Molecular Weight; D010455:Peptides / Q000737:chemistry; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D020543:Proteome / Q000032:analysis*; D020539:Sequence Analysis, Protein; D016622:Silver Staining; D013154:Spleen / Q000166:cytology
D010455:Peptides; D011506:Proteins; D020543:Proteome