Characterization of protein expression levels with label-free detected reverse phase protein arrays

Xuexue Guo; Yihong Deng; Chenggang Zhu; Junlong Cai; Xiangdong Zhu; James P Landry; Fengyun Zheng; Xunjia Cheng; Yiyan Fei

doi:10.1016/j.ab.2016.06.027

Characterization of protein expression levels with label-free detected reverse phase protein arrays

Anal Biochem. 2016 Sep 15:509:67-72. doi: 10.1016/j.ab.2016.06.027. Epub 2016 Jun 29.

Authors

Xuexue Guo¹, Yihong Deng², Chenggang Zhu¹, Junlong Cai², Xiangdong Zhu³, James P Landry³, Fengyun Zheng⁴, Xunjia Cheng², Yiyan Fei⁵

Affiliations

¹ Department of Optical Science and Engineering, Shanghai Engineering Research Center for Ultra-Precision Optical Manufacturing, Green Photoelectron Platform, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Fudan University, Shanghai, 200433, China.
² Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China.
³ Department of Physics, University of California, Davis, CA, 95616, USA.
⁴ Institutes of Biomedical Science, Fudan University, Shanghai, 200032, China.
⁵ Department of Optical Science and Engineering, Shanghai Engineering Research Center for Ultra-Precision Optical Manufacturing, Green Photoelectron Platform, Key Laboratory of Micro and Nano Photonic Structures (Ministry of Education), Fudan University, Shanghai, 200433, China. Electronic address: [email protected].

PMID: 27372609
DOI: 10.1016/j.ab.2016.06.027

Abstract

In reverse-phase protein arrays (RPPA), one immobilizes complex samples (e.g., cellular lysate, tissue lysate or serum etc.) on solid supports and performs parallel reactions of antibodies with immobilized protein targets from the complex samples. In this work, we describe a label-free detection of RPPA that enables quantification of RPPA data and thus facilitates comparison of studies performed on different samples and on different solid supports. We applied this detection platform to characterization of phosphoserine aminotransferase (PSAT) expression levels in Acanthamoeba lysates treated with artemether and the results were confirmed by Western blot studies.

Keywords: Label-free detection; Normalization; Quantification; Reverse phase protein arrays.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acanthamoeba castellanii / chemistry
Acanthamoeba castellanii / metabolism*
Protein Array Analysis / methods*
Protozoan Proteins / chemistry
Protozoan Proteins / metabolism*

Substances

Protozoan Proteins