Human Semaphorin 4A Antibody

  
  • Species Reactivity
    Human
  • Specificity
    Detects human Semaphorin 4A in direct ELISAs and Western blots. In direct ELISA, no cross-reactivity with recombinant human Semaphorin 4B, 4C, or 4D is observed and approximately 5% cross-reactivity with recombinant mouse Semaphorin 4A is observed.
  • Source
    Monoclonal Mouse IgG1 Clone # 741509
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Semaphorin 4A
    Gly32-His683
    Accession # Q9H3S1
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    2 µg/mL
    See below
  • Immunohistochemistry
    8-25 µg/mL
    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
Detection of Human Semaphorin 4A by Western Blot. Western blot shows lysates of human dendritic cells. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Semaphorin 4A Monoclonal Antibody (Catalog # MAB46941) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Semaphorin 4A at approximately 90-95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
Semaphorin 4A in Mouse Embryo. Semaphorin 4A was detected in immersion fixed frozen sections of mouse embryo (13 d.p.c.) using Mouse Anti-Human Semaphorin 4A Monoclonal Antibody (Catalog # MAB46941) at 25 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal processes. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.5 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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 4A
Semaphorin 4A (Sema4A, previously semB) is a Class 4 transmembrane Semaphorin with activity in the immune and nervous systems (1). The 761 amino acid (aa) human Sema4A precursor contains a 32 aa signal sequence, a 651 aa extracellular domain (ECD) containing sema, PSI and C2-type immunoglobulin domains, a 21 aa transmembrane domain, and a 57 aa cytoplasmic domain with two Ser/Thr phosphorylation sites (2). Human Sema4A ECD shares 87%, 87%, 86% and 85% aa identity with mouse, rat, bovine and canine Sema4A, respectively, and shares 32-37% aa identity with other human Sema4 family members. Of six reported splice variants with 723, 629, 370, 321, 236 and 220 aa, five lack the N-terminus and/or portions of the sema domain, and three lack the transmembrane and cytoplasmic domains in the C-terminus (3). Sema4A was first described as a molecule that enhances T cell activation and interacts with TIM-2 (T cell immunoglobulin and mucin domain-2 (4). Mice with targeted disruption of Sema4A show defects in dendritic cell-mediated T cell priming and Th1 responses (5). Roles for Sema4A have also been identified in the brain, the endothelium and the eye. It mediates hippocampal neuron growth cone collapse in vitro through interaction of the sema domain with plexin-B1 (6). Interaction of Sema4A with endothelial cell plexin-D1 causes opposition to the angiogenic, proliferative, chemotactic and integrin-mediated adhesive actions of VEGF (7). The retina of Sema4A-/- mice shows severe degeneration, and mutations of Sema4A are associated with retinitis pigmentosa and cone rod dystrophy in humans (8, 9).
  • References:
    1. Kumanogoh, A. et al. (2003) J. Cell Sci. 116:3463.
    2. Swissprot Accession # Q9H3S1.
    3. Entrez Accession # CAI15528, CAI15529, CAI15531, CAI15532, CAI15533 and EAW52993.
    4. Kumanogoh, A. et al. (2002) Nature 419:629.
    5. Kumanogoh, A. et al. (2005) Immunity 22:305.
    6. Yukawa, K. et al. (2005) Int. J. Mol. Med. 16:115.
    7. Toyofuku, T. et al. (2007) EMBO J. 26:1373.
    8. Rice, D.S. et al. (2004) Invest. Ophthalmol. Vis. Sci. 45:2767.
    9. Abid, A. et al. (2007) J. Med. Genet. 43:378.
  • Entrez Gene IDs:
    64218 (Human); 20351 (Mouse); 310630 (Rat)
  • Alternate Names:
    CORD10; RP35; Sema B; sema domain, immunoglobulin domain (Ig), transmembrane domain (TM) and shortcytoplasmic domain, (semaphorin) 4A; SEMA4A; SEMAB; Semaphorin 4A; semaphorin-4A; semaphorin-B; SEMB; SEMBFLJ12287
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