Human Semaphorin 3E APC-conjugated Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Semaphorin 3E in direct ELISAs. In direct ELISAs, approximately 15% cross-reactivity with recombinant human (rh) Semaphorin 3B is observed and no cross-reactivity with rhSemaphorin 6A is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 400513
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Semaphorin 3E
    Thr25-Ser775 (Arg557Ala and Arg560Ala)
    Accession # O15041
  • Formulation
    Supplied in a saline solution containing BSA and Sodium Azide.
  • Label
    Allophycocyanin
Applications
  •  
    Recommended
    Concentration
    Sample
  • Intracellular Staining by Flow Cytometry
    10 µL/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Intracellular Staining by Flow Cytometry
Detection of Semaphorin 3E in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes stimulated with PHA were stained with Rat Anti-Mouse CD3 PE‑conjugated Monoclonal Antibody (Catalog # FAB4841P) and either (A) Mouse Anti-Human Semaphorin 3E APC‑conjugated Monoclonal Antibody (Catalog # IC32391A) or (B) Mouse IgG1 Allophycocyanin Isotype Control (Catalog # IC002A). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Intracellular Staining by Flow Cytometry
Detection of Semaphorin 3E in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells stimulated with PHA were stained with Mouse Anti-Human Semaphorin 3E APC‑conjugated Monoclonal Antibody (Catalog # IC32391A, filled histogram) or isotype control antibody (Catalog # IC002A, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
  • Stability & Storage
    Protect from light. Do not freeze.
    • 12 months from date of receipt, 2 to 8 °C as supplied.
Background: Semaphorin 3E

Semaphorin 3E (Sema3E; previously SemaH) is a 90-95 kDa member of the Class 3 (secreted) semaphorins which, in human, share 40-50% amino acid (aa) sequence identity. Class 3 semaphorins are potent chemorepellents that function in axon guidance and/or vascular tip cell guidance during development (1). Sema3E is highly expressed in developing somites, where it acts as a repulsive cue for PlexinD1-expressing endothelial cells of adjacent intersomitic vessels (2, 3). Crystal structures of semaphorins reveal that the 500 aa N-terminal Sema domain forms a seven-blade beta -propeller similar to that found in integrin molecules. This is accompanied by 14 conserved cysteine residues and one or more N-glycosylation sites are thought critical for forming the secondary structure (4). C-terminal to the Sema domain, Sema3E has a consensus sequence for furin cleavage which, when used, creates a 61 kDa form that does not dimerize, and is highly expressed in tumor cell lines with metastatic potential (5, 6). Further C-terminal are a cysteine-knot plexin/semaphorin/integrin (PSI) domain, an Ig-like domain, a cysteine for dimerization and a basic domain containing another furin cleavage site. Dimerization and cleavage at the C-terminal site are required for repulsing activity of class 3 semaphorins (7). Human Sema3E shares 90%, 85% and 57% aa sequence identity with mouse, bovine and canine Sema3E, respectively. Like other semaphorins, Sema3E signaling is transduced by a transmembrane Plexin dimer, which also has a Sema domain and is coupled to kinase pathways. Unlike other Class 3 semaphorins, Sema3E binds directly to its plexin and does not require interaction with a neuropilin for activity (7). Genetic disruption of either Sema3E or PlexinD1 creates mouse mutants with excessive and disorganized vascular growth and branching, indicating the importance of this ligand-receptor pair for vascular guidance (3, 8).

  • References:
    1. Eichmann, A. et al. (2005) Genes Dev. 19:1013.
    2. Cohen, S. et al. (2005) Eur. J. Neurosci. 21:1767.
    3. Gu, C. et al. (2005) Science 307:265.
    4. Gherardi, E. et al. (2004) Curr. Opin. Struct. Biol. 14:669.
    5. Christensen, C. et al. (1998) Cancer Res. 58:1238.
    6. Christensen, C. et al. (2005) Cancer Res. 65:6167.
    7. Adams, R. H. et al. (1997) EMBO J. 16:6077.
    8. Gitler, A. D. et al. (2004) Dev. Cell 7:107.
  • Entrez Gene IDs:
    9723 (Human); 20349 (Mouse)
  • Alternate Names:
    (semaphorin) 3E; coll-5; KIAA0331M-SEMAH; sema domain, immunoglobulin domain (Ig), short basic domain, secreted; Sema3E; SEMAH; SEMAHM-SemaK; Semaphorin 3E; semaphorin-3E
Related Research Areas
Isotype Controls
Description Application Cat# Citations Images  

Mouse IgG1 APC-conjugated Antibody

Ctrl IC002A 8  
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Staining Reagents
Description Application Cat# Citations Images  

Flow Cytometry Permeabilization/Wash Buffer I (1X)

Flow FC005 5
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Flow Cytometry Fixation Buffer (1X)

Flow FC004 3
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Flow Cytometry Fixation & Permeabilization Buffer Kit I

Flow FC009 3
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Flow Cytometry Fixation/Permeabilization Buffer I (1X)

Flow FC007 2
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