dorsal/arxiv
View SchemaToward a better analysis of secreted proteins: the example of the myeloid cells secretome
| Authors | Mireille Chevallet, Hélène Diemer, Alain van Dorsselaer, Christian Villiers, Thierry Rabilloud |
|---|---|
| Categories | |
| ArXiv ID | q-bio/0703006 |
| URL | https://arxiv.org/abs/q-bio/0703006 |
Abstract
The analysis of secreted proteins represents a challenge for current proteomics techniques. Proteins are usually secreted at low concentrations in the culture media, which makes their recovery difficult. In addition, culture media are rich in salts and other compounds interfering with most proteomics techniques, which makes selective precipitation of proteins almost mandatory for a correct subsequent proteomics analysis. Last but not least, the non-secreted proteins liberated in the culture medium upon lysis of a few dead cells heavily contaminate the so-called secreted proteins preparations. Several techniques have been used in the past for concentration of proteins secreted in culture media. These techniques present several drawbacks, such as coprecipitation of salts or poor yields at low protein concentrations. Improved techniques based on carrier-assisted trichloroacetic acid precipitation are described and discussed in this paper. These techniques have been used to analyse the secretome of myeloid cells (macrophages, dendritic cells) and enabled to analyze proteins secreted at concentrations close to 1 ng/ml, thereby allowing to detect some of the cytokines (TNF, IL-12) secreted by the myeloid cells upon activation by bacterial products.
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"abstract": "The analysis of secreted proteins represents a challenge for current\nproteomics techniques. Proteins are usually secreted at low concentrations in\nthe culture media, which makes their recovery difficult. In addition, culture\nmedia are rich in salts and other compounds interfering with most proteomics\ntechniques, which makes selective precipitation of proteins almost mandatory\nfor a correct subsequent proteomics analysis. Last but not least, the\nnon-secreted proteins liberated in the culture medium upon lysis of a few dead\ncells heavily contaminate the so-called secreted proteins preparations. Several\ntechniques have been used in the past for concentration of proteins secreted in\nculture media. These techniques present several drawbacks, such as\ncoprecipitation of salts or poor yields at low protein concentrations. Improved\ntechniques based on carrier-assisted trichloroacetic acid precipitation are\ndescribed and discussed in this paper. These techniques have been used to\nanalyse the secretome of myeloid cells (macrophages, dendritic cells) and\nenabled to analyze proteins secreted at concentrations close to 1 ng/ml,\nthereby allowing to detect some of the cytokines (TNF, IL-12) secreted by the\nmyeloid cells upon activation by bacterial products.",
"arxiv_id": "q-bio/0703006",
"authors": [
"Mireille Chevallet",
"H\u00e9l\u00e8ne Diemer",
"Alain van Dorsselaer",
"Christian Villiers",
"Thierry Rabilloud"
],
"categories": [
"q-bio.GN"
],
"title": "Toward a better analysis of secreted proteins: the example of the myeloid cells secretome",
"url": "https://arxiv.org/abs/q-bio/0703006"
},
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