Human Semaphorin 3E Antibody

Catalog #: MAB32391 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB32391
MAB32391-SP

Detection of Semaphorin 3E in Human T Cells by Flow Cytometry.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Semaphorin 3E 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

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Intracellular Staining by Flow Cytometry

0.25 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 T Cells antibody by Flow Cytometry.

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Detection of Semaphorin 3E in Human T Cells by Flow Cytometry. Human T cells were treated for 3‑5 days with 1 µg/mL PHA then stained with Mouse Anti-Human Semaphorin 3E Monoclonal Antibody (Catalog # MAB32391, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

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

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.
Adams, R. H. et al. (1997) EMBO J. 16:6077.
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