Issue 23, 2012
From the journal:

Analyst

A CE–LIF method to monitor autophagy by directly detecting LC3 proteins in HeLa cells

Yue Jin,a   Cheng Chen,a   Lingchen Meng,a   Jitao Chen,a   Meixian Li,a   Zhiwei Zhu*a  and  Jian Lin*b  

* Corresponding authors

a Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, PR China
E-mail: zwzhu@pku.edu.cn
Fax: +86 10 62751708
Tel: +86 10 62757953

b Beijing Nuclear Magnet Resonance Center, Peking University, Beijing 100871, PR China
E-mail: linjian@pku.edu.cn
Fax: +86 10 62751708
Tel: +86 10 62755835

Abstract

A fast, simple and quantitative approach was established for monitoring autophagy in HeLa cells by directly detecting the conversion of green fluorescent protein (GFP) labelled autophagy markers, GFP-LC3-I to GFP-LC3-II, in crude cellular extract using capillary electrophoresis (CE) with laser-induced fluorescence (LIF). Compared with the traditional methods, this proposed method is simpler and more reliable. Moreover, high sensitivity, with a limit of detection (LOD) of 0.48 ppt (bovine serum albumin as standard protein), was obtained by an in-capillary derivatization method coupled with a field-enhanced sample injection (FESI) technique, this may allow the success of this technique in the detection of endogenous markers of autophagy in cells.

Graphical abstract: A CE–LIF method to monitor autophagy by directly detecting LC3 proteins in HeLa cells

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Article information

DOI
https://doi.org/10.1039/C2AN36011J
Article type
Paper
Submitted
25 Jul 2012
Accepted
20 Sep 2012
First published
25 Sep 2012

Analyst, 2012,137, 5571-5575

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A CE–LIF method to monitor autophagy by directly detecting LC3 proteins in HeLa cells

Y. Jin, C. Chen, L. Meng, J. Chen, M. Li, Z. Zhu and J. Lin, Analyst, 2012, 137, 5571 DOI: 10.1039/C2AN36011J

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