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Monoclonal Antibodies

Anti-Zebrafish DeltaC [zdc2]

Invented by Dr Gavin Wright from Wellcome Trust Sanger Institute
Invented at Cancer Research UK London Research Institute: Lincoln's Inn Fields
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Adapted from Giudicelli et al. 2007. PLoS Biol. 5(6):e150. PMID: 17535112. Figure. The Pattern of deltaC Transcripts Compared with the Pattern of DeltaC Protein in a Doubly Stained Specimen The left-hand panel shows the distribution of deltaC mRNA, as revealed by ISH; arrows indicate the three most recently formed somites. The middle panel shows the distribution of DeltaC protein in the same optical section, immunostained with the zdc2 monoclonal antibody. The right-hand panel shows the two patterns superimposed in the doubly stained specimen. The protein pattern is shifted by almost exactly one somite width relative to the mRNA pattern; one somite cycle time earlier, the cells currently displaying peak levels of protein would have been in the locations of the cells currently expressing the peak levels of mRNA and would have been expressing those levels of mRNA themselves. Thus there is a delay of approximately one somite cycle time (30 min) from the accumulation of the mRNA in the cell to the accumulation of the protein translated from it.
Adapted from Wright GJ, Giudicelli F, Soza-Ried C, Hanisch A, Ariza-McNaughton L, Lewis J. DeltaC and DeltaD interact as Notch ligands in the zebrafish segmentation clock. Development. 2011 Jul;138(14):2947-56. doi: 10.1242/dev.066654. Epub 2011 Jun 8. PMID: 21653612. Expression of DeltaC and DeltaD in retina and brain. (A-D) Sections of wild-type (WT) and mib mutant zebrafish retina at 24 hpf, immunostained with zdc2 and zdd2. (E,F) In situ hybridisation (ISH) of wild-type specimens (wholemounts) with probes detecting deltaC and deltaD mRNA. (G,H) Sections of hindbrain at 24 hpf, immunostained for DeltaC and DeltaD. (I-K) Section of retina and adjacent brain tissue in a 48 hpf embryo, doubly immunostained for DeltaC and DeltaD. In the retina the two proteins largely colocalise, but in the brain they are in distinct sets of cells. A-D and I-K are single confocal optical sections of 15 μm cryosections; G,H are projections of a small z-stack. A-D and G,H are counterstained for actin with fluorescent phalloidin (green).
Adapted from Wright GJ, Giudicelli F, Soza-Ried C, Hanisch A, Ariza-McNaughton L, Lewis J. DeltaC and DeltaD interact as Notch ligands in the zebrafish segmentation clock. Development. 2011 Jul;138(14):2947-56. doi: 10.1242/dev.066654. Epub 2011 Jun 8. PMID: 21653612. Selection of non-cross-reactive monoclonal antibodies that recognise formalin-treated zebrafish DeltaC and DeltaD glycoproteins. (A) Soluble recombinant Delta proteins used as immunogens, each comprising the extracellular region of zebrafish DeltaA, B, C or D, a C-terminal tag of rat Cd4 domains 3 and 4 (Cd4d3+4) and a short peptide that can be enzymatically monobiotinylated. (B) Selection of antibodies that recognise formalin-fixed DeltaD. (i) Untransfected (thin black line) or full-length DeltaD-transfected (thick blue line) HEK293T cells were stained with zdd2 monoclonal antibody and analysed by flow cytometry. (ii) The sensitivity of the zdd2 epitope to formalin fixation was assessed by flow cytometry of DeltaD-transfected cells fixed for 0 (black line), 40 (blue line) or 90 (red line) minutes before staining with zdd2. (C) Anti-zebrafish DeltaC and DeltaD monoclonal antibodies show little cross-reactivity within the zebrafish Delta family. The Cd4 antigenic tag and the four soluble Delta constructs were captured via their biotin tags to a streptavidin-coated ELISA plate before incubation with the indicated monoclonal antibodies. Antibody binding was quantified by a colorimetric assay using a phosphatase-conjugated secondary antibody. Bar chart shows mean ± s.d.; n=3. (D) Western blots of recombinant protein secreted from HEK293T cells transfected with tagged Delta constructs. R, reducing conditions; N, non-reducing conditions. All four antibodies reveal bands at the expected sizes for monomeric Delta and, more faintly, at approximately twice this size.
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Catalogue Number 151510
Applications ELISA FACS IHC IF WB
Antigen/Gene or Protein Targets Zebrafish DeltaC
Reactivity Zebrafish
Relevance Delta proteins activate Notch through a binding reaction that depends on their extracellular domains. During somitogenesis, Notch signalling via DeltaC keeps the oscillations of neighbouring cells in synchrony. DeltaC is also expressed in subsets of cells in the nervous system and elsewhere.
Host Mouse
Immunogen Extracellular region of zebrafish DeltaC protein
Positive Control HEK293 cells transfected with DeltaD and Zebrafish embryo cryosections (formulin fixed)
Subclass IgG2a
Myeloma Used Sp2/0-Ag14
Strain Balb/c
Research Area Aquaculture, Cell Cycle, Developmental Biology, Neurobiology

References

There are 6 reference entries for this reagent.

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References: 6 entries

Gallagher et al. 2017. Dev Biol. 429(1):225-239. PMID: 28648842.

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Quillien et al. 2014. Development. 141(7):1544-52. PMID: 24598161.

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Wright et al. 2011. Development. 138(14):2947-56. PMID: 21653612.

View  

DeltaC and DeltaD interact as Notch ligands in the zebrafish segmentation clock.

Europe PMC ID: 21653612

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Giudicelli et al. 2007. PLoS Biol. 5(6):e150. PMID: 17535112.

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Setting the tempo in development: an investigation of the zebrafish somite clock mechanism.

Europe PMC ID: 17535112

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References: 6 entries

Gallagher et al. 2017. Dev Biol. 429(1):225-239. PMID: 28648842.

View  

Quillien et al. 2014. Development. 141(7):1544-52. PMID: 24598161.

View  

Wright et al. 2011. Development. 138(14):2947-56. PMID: 21653612.

View  

DeltaC and DeltaD interact as Notch ligands in the zebrafish segmentation clock.

View  

Giudicelli et al. 2007. PLoS Biol. 5(6):e150. PMID: 17535112.

View  

Setting the tempo in development: an investigation of the zebrafish somite clock mechanism.

View  

Add a reference

Inventor Information