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Cq4 INVESTIGATOR Name Elisabeth Knust Address Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr. 108, Dresden 01307 Germany E-mail: [email protected] IMMUNOGEN Substance Name Origin Chemical Composition Developmental Stage IMMUNIZATION PROTOCOL Donor Animal Species Strain Sex Organ and tissue Immunization Dates immunized Amount of antigen Route of immunization Adjuvant FUSION Date Myeloma cell line Species Designation MONOCLONAL ANTIBODY Isotype Specificity Cell binding Immunohistology Antibody competition Species Specificity ANTIGEN Chemical properties Molecular weight Characterization Fusion protein between -galactosidase and amino acid 737-1703 of Drosophila Crumbs protein (cloned in pTRB vector). Expressed in bacteria, purified on PAA gel, eluted and precipitated by acetone. mouse BALB/c female 1990 1g intraperitoneal Freund’s complete, final boost: Freund’s incomplete 1991 mouse NS-1 IgG1, kappa light chain nd nd crossreacts with Drosophila virilis, not with mouse or C. elegans. transmembrane protein 234 kDa extracellular domain: 30 EGF-like repeats and 4 laminin-A G-domain like repeats cytoplasmic domain: 37 amino acids, highly conserved in homologous protein of C. elegans and human Immunoprecipitation Immunoblotting Purification Amino acid sequence analysis accession number: AAA28428 Functional effects loss of crumbs function prevents stabilization of the zonula adherens of epithelial cells, followed by the breakdown of the epithelial tissue structure and later cell death (whether Cq4 blocks this function has not been determined) Immunohistochemistry Crumbs protein is expressed in all epithelia of ectodermal origin in the Drosophila embryo from gastrulation onwards and in all imaginal discs and the follicle epithelium. It is restricted to a region apical to the zonula adherens. Good apical marker for epithelial cells. PUBLICATIONS : Tepass, U., Theres, C., and Knust, E. (1990). The Drosophila gene crumbs encodes an EGF-like protein, expressed on apical membranes of epithelial cells and required for organization of epithelia. Cell 61, 787-799. (Continued) Cq4 (Continued) Tepass, U., and Knust, E. (1990). Phenotypic and developmental analysis of mutations at the crumbs locus, a gene required for the development of epithelia in Drosophila melanogaster. Roux’s Arch. Dev. Biol. 199, 189-206. Tepass, U., and Knust, E. (1993). Crumbs and stardust act in a genetic pathway that controls the organization of epithelia in Drosophila melanogaster. Dev. Biol. 159, 311-326. Baumann, O. (2001). Posterior midgut epithelial cells differ in their organization of the membrane skeleton from other Drosophila epithelia. Exp. Cell Res. 270, 176-187. Morais-de-Sá, E., Mirouse, V., and St. Johnston, D. (2010). aPKC phosphorylation of Bazooka defines the apical/lateral border in Drosophila epithelial cells. Cell 141, 509-523. Maurel-Zaffran, C., Pradel, J., and Graba, Y. (2010). Reiterative use of signalling pathways controls multiple cellular events during Drosophila posterior spiracle organogenesis. Dev. Biol. 343, 18-27 Galy, A., Schenck, A., Sahin, H.B., Qurashi, A., Sahel, J.-A., Diebold, C., and Giangrande, A. (2011). CYFIP dependent Actin remodeling controls specific aspects of Drosophila eye morphogenesis. Dev. Biol. 359, 37-46. ACKNOWLEDGMENTS STATEMENT We have been asked by NICHD to ensure that all investigators include an acknowledgment in publications that benefit from the use of the DSHB's products. We suggest that the following statement be used: “The (select: hybridoma, monoclonal antibody, or protein capture reagent,) developed by [Investigator(s) or Institution] was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242.” Please send copies of all publications resulting from the use of Bank products to: Developmental Studies Hybridoma Bank Department of Biology The University of Iowa 028 Biology Building East Iowa City, IA 52242