Human Semaphorin 3E Alexa Fluor® 488-conjugated Antibody

Catalog # Availability Size / Price Qty
IC32391G-100UG
Detection of Semaphorin 3E in Human PBMCs by Flow Cytometry.
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Human Semaphorin 3E Alexa Fluor® 488-conjugated Antibody Summary

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 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 488 (Excitation= 488 nm, Emission= 515-545 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
0.25-1 µg/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.

Scientific Data

Intracellular Staining by Flow Cytometry Detection of Semaphorin 3E antibody in Human PBMCs antibody by Flow Cytometry. View Larger

Detection of Semaphorin 3E in Human PBMCs by Flow Cytometry. Human peripheral blood mononuclear cells (PBMCs) treated with PHA for were stained with Mouse Anti-Human CD3e APC-conjugated Monoclonal Antibody (Catalog # FAB100A) and either (A) Mouse Anti-Human Semaphorin 3E Alexa Fluor® 488-conjugated Monoclonal Antibody (Catalog # IC32391G) or (B) Mouse IgG1Alexa Fluor 488 Isotype Control (Catalog # IC002G). 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 antibody in Mouse Splenocytes antibody by Flow Cytometry. View Larger

Detection of Semaphorin 3E in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes treated with PHA were stained with Rat Anti-Mouse CD3 APC-conjugated Monoclonal Antibody (Catalog # FAB4841A) and either (A) Mouse Anti-Human Semaphorin 3E Alexa Fluor® 488-conjugated Monoclonal Antibody (Catalog # IC32391G) or (B) Mouse IgG1Alexa Fluor 488 Isotype Control (Catalog # IC002G). 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.

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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 one of six Class 3 (secreted) semaphorins which in the human share 40-50% amino acid (aa) 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 by a subset of motor neurons 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 b-propeller similar to that found in integrin molecules; 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 61kDa 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 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 identity with mouse, cow and dog 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) Developmental 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

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Product Specific Notices


This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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