Proteomics

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Proteome profiling of HaCaT keratinocytes


ABSTRACT: An important step in the proteomic analysis is the optimal extraction and the processing of protein material in order to ensure the highest sensitivity in downstream protein detection. Cell lysis is important for proteomic profiling with an eventual aim to discover differently expressed proteins (DEPs). Herein for cell lysis, two sample preparation protocols were compared: 0.2% SDS-based solubilization combined with the 1DE-gel concentration (Protocol 1) and osmotic shock (Protocol 2). HaCaT keratinocytes were cultured with/without sodium dodecyl sulfate (SDS) and used to test the hypothesis that SDS might be associated with the initiation and development of malignancies. Protocol 1 increased sensitivity and coverage of HaCaT proteome, it permitted us to reveal the provided new insights into the skin impact of SDS (widely used and rather non-toxic component of household chemicals, cosmetic products, shampoos and so on). Our results suggest the possible role of SDS in skin cancerogenesis.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Keratinocyte

DISEASE(S): Disease Free

SUBMITTER: Timur Shkrigunov  

LAB HEAD: Natalia Petushkova

PROVIDER: PXD035202 | Pride | 2022-07-20

REPOSITORIES: Pride

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Publications

Protocol for Increasing the Sensitivity of MS-Based Protein Detection in Human Chorionic Villi.

Shkrigunov Timur T   Pogodin Pavel P   Zgoda Victor V   Larina Olesya O   Kisrieva Yulia Y   Klimenko Maria M   Latyshkevich Oleg O   Klimenko Peter P   Lisitsa Andrey A   Petushkova Natalia N  

Current issues in molecular biology 20220509 5


An important step in the proteomic analysis of missing proteins is the use of a wide range of tissues, optimal extraction, and the processing of protein material in order to ensure the highest sensitivity in downstream protein detection. This work describes a purification protocol for identifying low-abundance proteins in human chorionic villi using the proposed "1DE-gel concentration" method. This involves the removal of SDS in a short electrophoresis run in a stacking gel without protein separ  ...[more]

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