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DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
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Related Experiment Video

Updated: Sep 17, 2025

A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue
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Optimized protein extraction protocol from human skin samples.

Ana Paula Carvalho Reis1, Giovanna Azevedo Celestrino1, Talita Souza Siqueira2

  • 1Laboratory of Medical Mycology LIM-53, Division of Clinical Dermatology, Instituto de Medicina Tropical, Hospital das Clínicas, Faculty of Medicine, Universidade de São Paulo, Avenida Doutor Enéas de Carvalho Aguiar, 470, São Paulo, SP, 05403-000, Brazil.

Biology Methods & Protocols
|June 27, 2025
PubMed
Summary
This summary is machine-generated.

Researchers optimized a human skin protein extraction method, identifying ~6000 skin proteins. This advance aids understanding of skin biology and disease biomarker discovery.

Keywords:
extraction of proteinsmass spectrometryproteomicsprotocolskin

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Area of Science:

  • Proteomics
  • Dermatology
  • Biochemistry

Background:

  • The skin, the body's largest organ, harbors numerous diseases.
  • Limited data exists on skin proteomics due to tissue complexity.
  • Understanding skin protein profiles is crucial for disease research.

Purpose of the Study:

  • To develop and optimize a robust protocol for human skin protein extraction.
  • To enhance the identification of proteins in healthy and diseased skin.
  • To facilitate the discovery of novel biomarkers and therapeutic targets for skin conditions.

Main Methods:

  • Adapted protocol using chemical and mechanical lysis (SDS, TEAB, inhibitor cocktail, FastPrep-24 5G).
  • Protein purification and digestion via Single-pot, solid phase, sample preparation (SP3) method.
  • Analysis using nano liquid chromatography coupled with tandem mass spectrometry (nLC-MS/MS).

Main Results:

  • Successfully identified approximately 6000 proteins from human skin samples.
  • Quantified proteins from both healthy individuals and patients with sporotrichosis.
  • Demonstrated the efficacy of the optimized extraction and analysis workflow.

Conclusions:

  • The improved methodology significantly enhances human skin protein identification.
  • This approach offers new perspectives for skin biology research.
  • Potential for discovering biomarkers and therapeutic targets for cutaneous diseases.