Hydrogel Pore-Size Modulation for Enhanced Single-Cell Western Blotting

Adv Mater. 2016 Jan 13;28(2):327-334. doi: 10.1002/adma.201503939. Epub 2015 Nov 16.

Abstract

Pore-gradient microgel arrays enable thousands of parallel high-resolution single-cell protein electrophoresis separations for targets accross a wide molecular mass (25-289 kDa), yet within 1 mm separation distances. Dual crosslinked hydrogels facilitate gel-pore expansion after electrophoresis for efficient and uniform immunoprobing. The photopatterned, light-activated, and acid-expandable hydrogel underpins single-cell protein analysis, here for oncoprotein-related signaling in human breast biopsy.

Keywords: cancer therapies; gradient gels; grayscale masks; hydrogel pore expansion; western blots.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Blotting, Western / instrumentation*
  • Blotting, Western / methods
  • Breast Neoplasms / metabolism
  • Cell Line, Tumor
  • Electrophoresis, Polyacrylamide Gel / instrumentation
  • Electrophoresis, Polyacrylamide Gel / methods
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Hydrogels* / chemistry
  • Hydrogels* / radiation effects
  • Light
  • Nucleocytoplasmic Transport Proteins / metabolism
  • Porosity
  • Receptor, ErbB-2 / metabolism
  • Single-Cell Analysis / instrumentation*
  • Single-Cell Analysis / methods
  • TOR Serine-Threonine Kinases / metabolism

Substances

  • EIF4ENIF1 protein, human
  • Hydrogels
  • Nucleocytoplasmic Transport Proteins
  • Green Fluorescent Proteins
  • MTOR protein, human
  • ERBB2 protein, human
  • Receptor, ErbB-2
  • TOR Serine-Threonine Kinases
  • Extracellular Signal-Regulated MAP Kinases