Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 859, Issue 1, Pages 30-36Publisher
ELSEVIER
DOI: 10.1016/j.jchromb.2007.09.022
Keywords
phosphoamino acids; beta-cyclodextrin; capillary electrophoresis; naphthalene-2,3-dicarboxaldehyde
Ask authors/readers for more resources
A rapid and sensitive capillary electrophoresis (CE) method coupled with fluorescence detection was developed for identification of protein phosphorylation by determination of phosphoamino acids. Naphthalene-2,3-dicarboxaldehycle (NDA), a fluorescence derivatization reagent, was used to label protein hydrolysate. The optimal derivatization reaction was performed with 3.5 mM NDA, 40 mM NaCN and 20 mM borate buffer (pH 10.0) for 15 min. The baseline separation of three phosphorylated amino acids could be obtained in less than 180 s with good repeatability by using 30 mM borate (pH 9.2) containing 2.0 mM P-cyclodextrin (P-CD) as the running; buffer. The detection limits for phosphothreonine, phosphotyrosine and phosphoserine were 7.0 x 10(-9) M, 5.6 x 10(-9) M and 7.2 x 10(-9) M, respectively (S/N = 3). Also, the interference from other protein amino acids with large molar excess over that of phosphoamino acids was studied. With beta-casein as the analysis protein, this method was successfully validated. (C) 2007 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available