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.
Monoclonal Mouse IgG1 Clone # 400513
Protein A or G purified from hybridoma culture supernatant
Mouse myeloma cell line NS0-derived recombinant human Semaphorin 3E Thr25-Ser775 (Arg557Ala and Arg560Ala) Accession # O15041
Supplied in a saline solution containing BSA and Sodium Azide.
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.
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
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).
Eichmann, A. et al. (2005) Genes Dev. 19:1013.
Cohen, S. et al. (2005) Eur. J. Neurosci. 21:1767.
Gu, C. et al. (2005) Science 307:265.
Gherardi, E. et al. (2004) Curr. Opin. Struct. Biol. 14:669.
Christensen, C. et al. (1998) Cancer Res. 58:1238.
Christensen, C. et al. (2005) Cancer Res. 65:6167.
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