title: Hydrogel Pore-Size Modulation for Enhanced Single-Cell Western Blotting.
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.
authors.affiliation: Department of Bioengineering, University of California Berkeley, Berkeley, California, CA 94720, USA.
authors.forename: Todd A
authors.initials: TA
authors.lastname: Duncombe
authors.affiliation: Department of Bioengineering, University of California Berkeley, Berkeley, California, CA 94720, USA.
authors.forename: Chi-Chih
authors.initials: CC
authors.lastname: Kang
authors.affiliation: Department of Bioengineering, University of California Berkeley, Berkeley, California, CA 94720, USA.
authors.forename: Santanu
authors.initials: S
authors.lastname: Maity
authors.affiliation: Division of Oncology, Department of Medicine, Stanford University, Stanford, California, CA 94305, USA.
authors.forename: Toby M
authors.initials: TM
authors.lastname: Ward
authors.affiliation: Division of Oncology, Department of Medicine, Stanford University, Stanford, California, CA 94305, USA.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Pegram
authors.affiliation: Department of Bioengineering, University of California Berkeley, Berkeley, California, CA 94720, USA.
authors.forename: Niren
authors.initials: N
authors.lastname: Murthy
authors.affiliation: Department of Bioengineering, University of California Berkeley, Berkeley, California, CA 94720, USA.
authors.forename: Amy E
authors.initials: AE
authors.lastname: Herr
mesh_terms: D015153:Blotting, Western / Q000295:instrumentation* / Q000379:methods; D001943:Breast Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020100:Hydrogels / Q000737:chemistry / Q000528:radiation effects; D008027:Light; D029543:Nucleocytoplasmic Transport Proteins / Q000378:metabolism; D016062:Porosity; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism
chemical_list: D048049:Extracellular Signal-Regulated MAP Kinases; D049452:Green Fluorescent Proteins; D020100:Hydrogels; D029543:Nucleocytoplasmic Transport Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D058570:TOR Serine-Threonine Kinases; C411571:EIF4ENIF1 protein, human; C546842:MTOR protein, human; C508053:ERBB2 protein, human
keywords: cancer therapies; gradient gels; grayscale masks; hydrogel pore expansion; western blots