Human Neuropilin‑1 Antibody

3 citations
  • Specificity
    Detects human Neuropilin-1 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant rat Neuropilin-1 is observed, and less than 1% cross-reactivity with recombinant human Neuropilin-2 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Neuropilin‑1
    Accession # NP_001019799
  • 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.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
  • Western Blot
    1 µg/mL
    See below
  • Flow Cytometry
    2.5 µg/106 cells
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Blockade of Receptor-ligand Interaction
    0.25-1 µg/mL
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 Neuropilin‑1 by Western Blot. Western blot shows lysates of MDA‑MB‑231 human breast cancer cell line and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Neuropilin‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3870) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Neuropilin‑1 at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Neuropilin-1 in Human Pancreatic Cancer Tissue. Neuropilin-1 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Sheep Anti-Human Neuropilin-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3870) at 10 µg/mL overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to the cytoplasm and plasma membrane of cancer cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Neuropilin‑1 in HUVEC Human Cells by Flow Cytometry. HUVEC human umbilical vein endothelial cells were stained with Sheep Anti-Human Neuropilin‑1 Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF3870, filled histogram) or control antibody (Catalog # 5-001-A, open histogram), followed by NorthernLights™ 637-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL011).
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • 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: Neuropilin-1

Neuropilin-1 (Npn-1, previously neuropilin; also CD304/BDCA4) is a 130-140 kDa type I transmembrane (TM) glycoprotein that regulates axon guidance and angiogenesis (1-4). The full-length 923 amino acid (aa) human Npn-1 contains a 623 aa extracellular domain (ECD) that shows 92-95% aa identity with mouse, rat, bovine, and canine Npn-1 (3, 4). The ECD contains two N-terminal CUB domains (termed a1a2), two domains with homology to coagulation factors V and VIII (b1b2) and a MAM (meprin) domain (c). C-terminally divergent splice variants with 704, 644, 609, and 551 aa lack the MAM and TM domains and are demonstrated or presumed to be soluble antagonists (1, 5-7). A 906 aa form lacks a TM segment, but secretion has not been found (8). The sema domains of Class III secreted semaphorins such as Sema3A bind Npn-1 a1a2 (9). Heparin, the heparin-binding forms of VEGF (VEGF165, VEGF-B, and VEGF-E), PlGF (PlGF‑2), and the C-terminus of Sema3 bind the b1b2 region (9, 10). Npn-1 and Npn-2 share 48% aa identity within the ECD and can form homo- and hetero-oligomers via interaction of their MAM domains (1). Neuropilins show partially overlapping expression in neuronal and endothelial cells during development (1, 2). Both neuropilins act as co-receptors with plexins, mainly plexin A3 and A4, to bind class III semaphorins that mediate axon repulsion (11). However, only Npn-1 binds Sema3A, and only Npn-2 binds Sema3F (1). Both are co-receptors with VEGF R2 (also called KDR or Flk-1) for VEGF165 binding (1). Sema3A signaling can be blocked by VEGF165, which has higher affinity for Npn-1 (12). Npn-1 is preferentially expressed in arteries during development or those undergoing remodeling (1, 2). Npn-1 is also expressed on dendritic cells and mediates DC-induced T cell proliferation (13).

  • References:
    1. Bielenberg, D.R. et al. (2006) Exp. Cell Res. 312:584. 
    2. Gu, C. et al. (2003) Dev. Cell 5:45. 
    3. He, Z. and M. Tessier-Lavigne (1997) Cell 90:739.
    4. Soker, S. et al. (1998) Cell 92:735. 
    5. Gagnon, M.L. et al. (2000) Proc. Natl. Acad. Sci. USA 97:2573. 
    6. Cackowski, F.C. et al. (2004) Genomics 84:82. 
    7. Rossignol, M. et al. (2000) Genomics 70:211.
    8. Tao, Q. et al. (2003) Angiogenesis 6:39.
    9. Gu, C. et al. (2002) J. Biol. Chem. 277:18069.
    10. Mamluk, R. et al. (2002) J. Biol. Chem. 277:24818.
    11. Yaron, A. et al. (2005) Neuron 45:513.
    12. Narazaki, M. and G. Tosato (2006) Blood 107:3892.
    13. Tordjman, R. et al. (2002) Nat. Immunol. 3477.
  • Alternate Names:
    BDCA-4; CD304; NRP1; BDCA4; CD304 antigen; DKFZp686A03134; DKFZp781F1414; neuropilin 1; neuropilin-1; NRPNP1; transmembrane receptor; Vascular endothelial cell growth factor 165 receptor; VEGF165RCD304
Related Research Areas

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Sample Type
  1. Neuropilin-1 mediates PDGF stimulation of vascular smooth muscle cell migration and signalling via p130Cas.
    Authors: Pellet-Many C, Frankel P, Evans IM, Herzog B, Junemann-Ramirez M, Zachary IC
    Biochem. J., 2011;435(3):609-18.
    Species: Human
    Sample Type: Cell Lysates
    Application: Western Blot,
  2. Alphav beta3 integrin limits the contribution of neuropilin-1 to vascular endothelial growth factor-induced angiogenesis.
    Authors: Robinson SD, Reynolds LE, Kostourou V, Reynolds AR, da Silva RG, Tavora B, Baker M, Marshall JF, Hodivala-Dilke KM
    J. Biol. Chem., 2009;284(49):33966-81.
    Species: Human
    Sample Type: Cell Lysates
    Application: Western Blot,
  3. Instruction of circulating endothelial progenitors in vitro towards specialized blood-brain barrier and arterial phenotypes.
    Authors: Boyer-Di Ponio J, El-Ayoubi F, Glacial F, Ganeshamoorthy K, Driancourt C, Godet M, Perriere N, Guillevic O, Couraud P, Uzan G
    PLoS ONE, 2014;9(1):e84179.
    Species: Mouse
    Sample Type: Whole Cells
    Application: ICC Fluorescence,


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