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| A Novel Purification Method for Lysosomes by Lmmunoprecipitation |
| ZHANG Lei, YANG Yu, WU Chuan-fang |
| Sichuan Functional Genome Laboratory, college of Life Science, Sichuan University, Chengdu 610064, China |
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Abstract A novel purification method for lysosomes by IP lysosomal membrane protein was explored in this paper. LAMP2-Flag was ligated into the eukaryotic expression vector pcDNA3.1, and was transfected into Hela cell line. The lysosomal location of LAMP2-Flag was conformed by Immunofluorescence labeling. Hela cell lines that can stably express LAMP2-Flag were cultured in large numbers. Highly pure lysosomes were isolated from cell extractions using anti-Flag immunoprecipitation under native condition. The purifiedlysosomes contained highlyenriched lysosomal marker proteins and enzymems, and no mitochondrial contamination. Our rResults indicate the efficiency of this method in the isolation of lysosomes of high purity from LAMP2-Flag expressing Hela cell lines.
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Received: 23 September 2013
Published: 25 November 2013
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