Human Neuropilin-1 Antibody

Catalog # Availability Size / Price Qty
AF3870
AF3870-SP
Detection of Human Neuropilin‑1 by Western Blot.
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Product Details
Citations (5)
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Human Neuropilin-1 Antibody Summary

Species Reactivity
Human
Specificity
Detects human Neuropilin-1 in direct ELISAs and Western blots. In direct ELISAs, less than 50% cross-reactivity with recombinant rat Neuropilin-1 and recombinant mouse 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
Phe22-Lys644
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
Unconjugated

Applications

Recommended Concentration
Sample
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
In a functional ELISA, 0.25-1 µg/mL of this antibody will block 50% of the binding of 20 ng/mL of Recombinant Human VEGF (Catalog # 293-VE) to immobilized Recombinant Human Neuropilin-1
(Catalog # 3870‑N1) coated at 0.75 µg/mL (100 µL/well). At 5 µg/mL, this antibody will block >90% of the binding.
 
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 

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

Western Blot Detection of Human Neuropilin‑1 by Western Blot. View Larger

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.

Flow Cytometry Detection of Neuropilin‑1 in HUVEC Human Cells by Flow Cytometry. View Larger

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).

Immunohistochemistry Neuropilin-1 in Human Pancreatic Cancer Tissue. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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.
Entrez Gene IDs
8829 (Human); 18186 (Mouse); 246331 (Rat)
Alternate Names
BDCA4; BDCA-4; CD304 antigen; CD304; DKFZp686A03134; DKFZp781F1414; neuropilin 1; Neuropilin1; Neuropilin-1; NRP1; NRPNP1; transmembrane receptor; Vascular endothelial cell growth factor 165 receptor; VEGF165RCD304

Product Datasheets

Citations for Human Neuropilin-1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

5 Citations: Showing 1 - 5
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  1. Neuropilin-1 mediates Neutrophil Elastase uptake and cross-presentation in breast cancer cells
    Authors: C Kerros, SC Tripathi, D Zha, JM Mehrens, A Sergeeva, AV Philips, N Qiao, HL Peters, H Katayama, P Sukhumalch, KE Ruisaard, AA Perakis, LS St John, S Lu, EA Mittendorf, K Clise-Dwye, AC Herrmann, G Alatrash, C Toniatti, SM Hanash, Q Ma, JJ Molldrem
    J. Biol. Chem., 2017;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  2. Conformational remodeling of the fibronectin matrix selectively regulates VEGF signaling.
    Authors: Ambesi A, McKeown-Longo P
    J Cell Sci, 2014;127(0):3805-16.
    Species: Human
    Sample Types: Whole Cells
    Applications: WB
  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 Types: Whole Cells
    Applications: ICC Fluorescence
  4. 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 Types: Cell Lysates
    Applications: WB
  5. 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 Types: Cell Lysates
    Applications: WB

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Human Neuropilin‑1 Antibody
By Anonymous on 04/21/2017