Cell kinase activity assay based on surface enhanced Raman spectroscopy.
Kinases control many important aspects of cell behavior, such as signal transduction, growth/differentiation, and tumorogenesis. Current methods for assessing kinase activity often require specific antibodies, and/or radioactive labeling. Here we demonstrated a novel detection method to assess kinase activity based on surface enhanced Raman spectroscopy (SERS). Raman signal was obtained after amplification by silver nanoparticles. The sensitivity of this method was comparable to fluorescence measurement of peptide concentration. When purified kinase enzyme was used, the detection limit was comparable to conventional radio-labeling method. We further demonstrated the feasibility to measure kinase activity in crude cell lysate. We suggested this SERS-based kinase activity assay could be a new tool for biomedical research and application.
Yue
Zhicao
Z
Department of Mechanical and Aerospace Engineering, Eng IV 44-121, 420 Westwood Plaza, University of California, Los Angeles, CA 90095, United States. zyue@ucla.edu
Zhuang
Fengfeng
F
Kumar
Rajar
R
Wong
Ieong
I
Cronin
Stephen B
SB
Liu
Yi-Hsin
YH
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002460:Cell Line; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D053616:Janus Kinase 3 / Q000737:chemistry / Q000378:metabolism; D053768:Metal Nanoparticles / Q000737:chemistry; D051379:Mice; D018625:Microscopy, Atomic Force; D010455:Peptides / Q000737:chemistry; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000737:chemistry / Q000378:metabolism; D012834:Silver / Q000737:chemistry; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013499:Surface Properties
D005456:Fluorescent Dyes; D010455:Peptides; D012834:Silver; D011494:Protein Kinases; D053616:Janus Kinase 3; D051057:Proto-Oncogene Proteins c-akt; D016650:Fluorescein-5-isothiocyanate