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.
Polyclonal Sheep IgG
Mouse myeloma cell line NS0-derived recombinant human Neuropilin‑1 Phe22-Lys644 Accession # NP_001019799
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.
<0.10 EU per 1 μg of the antibody by the LAL method.
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.
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.
are available in the Technical Information section on our website.
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.
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).
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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
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.
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).
Bielenberg, D.R. et al. (2006) Exp. Cell Res. 312:584.
Gu, C. et al. (2003) Dev. Cell 5:45.
He, Z. and M. Tessier-Lavigne (1997) Cell 90:739.
Soker, S. et al. (1998) Cell 92:735.
Gagnon, M.L. et al. (2000) Proc. Natl. Acad. Sci. USA 97:2573.
Cackowski, F.C. et al. (2004) Genomics 84:82.
Rossignol, M. et al. (2000) Genomics 70:211.
Tao, Q. et al. (2003) Angiogenesis 6:39.
Gu, C. et al. (2002) J. Biol. Chem. 277:18069.
Mamluk, R. et al. (2002) J. Biol. Chem. 277:24818.
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.
Submit a review and receive a $25US/€18/£15/$25CAN amazon gift card if you
include an image - $10US/€7/£6/$10CAN Amazon card for reviews without an image.
Limited to verified customers in USA, Canada and Europe.
Read what people are saying who have used Human Neuropilin-1 Antibody.