Separation of proteins by sodium dodecylsulfate capillary electrophoresis in hydroxypropylcellulose sieving matrix with laser-induced fluorescence detection.
Sodium dodecyl sulfate capillary electrophoresis by using hydroxypropylcellulose as the sieving matrix was developed for separation of proteins. 3-(2-furoyl)quinoline-2-carboxaldehyde, a fluorogenic dye, was used as the pre-column reagent to label proteins, which allows the use of laser-induced fluorescence to improve the detection sensitivity. Five standard proteins within the molecular mass range of 14,000-97,000 were used to test this method and a calibration curve was obtained between the molecular mass of these proteins and their peak migration times. This method was also applied to the separation of proteins from HT29 human colon adenocarcinoma cell extracts, and, typically, nearly 30 protein components could be resolved in a 20-min separation. Similar separation patterns were observed for the cell extract proteins when three running buffer systems were employed, indicating that buffer composition did not have much influence on the separation based on HPC sieving.
Hu
S
S
Department of Chemistry, University of Alberta, Edmonton, Canada.
Zhang
Z
Z
Cook
L M
LM
Carpenter
E J
EJ
Dovichi
N J
NJ
D002482:Cellulose / Q000031:analogs & derivatives* / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D007834:Lasers; D011506:Proteins / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D011506:Proteins; D012967:Sodium Dodecyl Sulfate; D002482:Cellulose; C008079:hydroxypropylcellulose