title: Ultrasensitive Protein Detection on Dot Blots and Western Blots with Semiconducting Polymer Dots.
abstract: We demonstrate ultrasensitive fluorescence imaging of proteins on dot blots and western blots using a bright, compact, and orange-emitting semiconducting polymer dot (CNPPV). We achieved a detection limit at the single-picogram level in dot blots; with conventional western blot, we detected 50 pg of transferrin and trypsin inhibitor after SDS-PAGE and transfer onto a PVDF membrane. In this protocol, we describe the preparation of Pdots and its streptavidin bioconjugate and the procedures of dot blot and western blots.
authors.affiliation: Department of Chemistry, University of Washington, 36 Bagley Hall, Seattle, WA, 98195, USA.
authors.forename: Fangmao
authors.initials: F
authors.lastname: Ye
authors.forename: Polina B
authors.initials: PB
authors.lastname: Smith
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Chiu
mesh_terms: D000818:Animals; D000906:Antibodies; D019921:Biotinylation; D015153:Blotting, Western / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D061848:Optical Imaging / Q000379:methods; D011108:Polymers / Q000737:chemistry*; D011506:Proteins / Q000032:analysis*; D045663:Quantum Dots / Q000737:chemistry*; D012666:Semiconductors; D019809:Streptavidin / Q000737:chemistry; D014168:Transferrin / Q000032:analysis
chemical_list: D000906:Antibodies; D005456:Fluorescent Dyes; D011108:Polymers; D011506:Proteins; D014168:Transferrin; D019809:Streptavidin