title: Generating a battery of monoclonal antibodies against native green fluorescent protein for immunostaining, FACS, IP, and ChIP using a unique adjuvant.
abstract: Using a unique, nontoxic adjuvant compound of poly(I:C) and anti-CD40 MAb, a battery of eight mouse monoclonal antibodies was generated against native green fluorescent protein. All were effective to varying degrees for immunostaining paraformaldehyde-fixed cells, six for staining sections of paraffin-embedded tissue, all to varying degrees in fluorescent-activated cell sorting, five for immunoprecipitation, and seven for chromatin immunoprecipitation. None worked in denaturing Western blots since the target was the native GFP protein. Both the hybridomas and antibodies are available at cost through DSHB, a non-profit National Resource created by the National Institutes of Health.
authors.affiliation: Developmental Studies Hybridoma Bank, The University of Iowa , Iowa City, Iowa.
authors.forename: Philip
authors.initials: P
authors.lastname: Sanchez
authors.forename: Karla J
authors.initials: KJ
authors.lastname: Daniels
authors.forename: Yang-Nim
authors.initials: YN
authors.lastname: Park
authors.forename: David R
authors.initials: DR
authors.lastname: Soll
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D030801:Animals, Genetically Modified; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis / Q000737:chemistry*; D015703:Antigens, CD; D015820:Cadherins / Q000276:immunology / Q000378:metabolism; D002176:Candida albicans / Q000235:genetics; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D005260:Female; D005434:Flow Cytometry; D019084:Fluorescent Antibody Technique, Indirect; D005656:Fungal Proteins / Q000276:immunology / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000276:immunology; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D011070:Poly I-C / Q000494:pharmacology*; D011485:Protein Binding; D011489:Protein Denaturation; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology*; D014157:Transcription Factors / Q000276:immunology / Q000378:metabolism; D015027:Zebrafish / Q000235:genetics
chemical_list: D000276:Adjuvants, Immunologic; D058846:Antibodies, Monoclonal, Murine-Derived; D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D011070:Poly I-C
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