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Workflow for proteomic analysis of purified lysosomes in cells lacking GRN v3
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Lysosomes are a major degradative organelle within eukaryotic cells. Previous work has developed a method wherein the TMEM192 protein is tagged on its C-terminus with an epitope tag in order to immunopurify (IP) lysosomes from cell extracts.1 This process is referred to as Lyso-IP. Such lysosomes can be used for proteomic analysis or for metabolomic analysis. The Lyso-IP is adapted from a previous reported method (Wyant et al., 2018). Here we also describe processing steps using proteomics after lysosome purification in the context of HeLa cells lacking the GRN gene. Such cells can be produced using the following protocol: DOI: dx.doi.org/10.17504/protocols.io.4r3l2oxqqv1y/v1.
Title: Workflow for proteomic analysis of purified lysosomes in cells lacking GRN v3
Description:
Lysosomes are a major degradative organelle within eukaryotic cells.
Previous work has developed a method wherein the TMEM192 protein is tagged on its C-terminus with an epitope tag in order to immunopurify (IP) lysosomes from cell extracts.
1 This process is referred to as Lyso-IP.
Such lysosomes can be used for proteomic analysis or for metabolomic analysis.
The Lyso-IP is adapted from a previous reported method (Wyant et al.
, 2018).
Here we also describe processing steps using proteomics after lysosome purification in the context of HeLa cells lacking the GRN gene.
Such cells can be produced using the following protocol: DOI: dx.
doi.
org/10.
17504/protocols.
io.
4r3l2oxqqv1y/v1.
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