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 (13)
FAQs
Supplemental Products
Reviews (5)

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 either lyophilized or 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.
 
Immunocytochemistry
5-15 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.

Scientific Data

Western Blot Detection of Human Neuropilin-1 antibody 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 antibody in HUVEC Human Cells antibody 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).

Immunocytochemistry Neuropilin-1 antibody in PC-3 and HLDM-2 Human Cell Lines by Immunocytochemistry (ICC). View Larger

Neuropilin‑1 in PC-3 and HLDM-2 Human Cell Lines. Neuropilin-1 was detected in immersion fixed PC-3 human prostate cancer cell line (positive staining) and HLDM-2 human Hodgkin's lymphoma cell line (negative staining) using Sheep Anti-Human Neuropilin-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3870) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry Neuropilin-1 antibody in Human Pancreatic Cancer Tissue by Immunohistochemistry (IHC-P). 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.

Knockdown Validated Detection of Human Neuropilin-1 by Knockdown Validated View Larger

Detection of Human Neuropilin-1 by Knockdown Validated NRP1-dependent EGFR/ERK signalling pathways are activated by EBV and are also associated with the promotion of EBV infection.(a,b) EBV infection activated EGFR/AKT and EGFR/ERK pathways. Serum-starved HNE1 cells were infected with EBV (a) or purified EBV (b) at an MOI of 5 × 103 for the indicated times. (c) EBV-activated EGFR/AKT or EGFR/ERK pathways were partially mediated by NRP1. After transfected with the indicated siRNA duplexes for 48 h, HNE1 cells were serum-starved and infected with EBV at an MOI of 5 × 103 for 1 h, followed by western blotting. For a–c, the intensity of the bands, determined with the ImageJ software, was shown as indicated. alpha -Tubulin, internal control. (d) Schematic diagram of RTKs (EGFR and VEGFR2)/NRP1/Ras/ERK and RTKs/NRP1/PI3K/AKT signalling cascade inhibitors targeting the indicated members of the pathways are shown. (e) EGFR/Ras/ERK signalling pathway was required for EBV infection. EGF-treated HNE1 and NPEC1-Bmi1 cells pre-incubated with Gefitinib (20 μM), Sorafenib (HNE1, 20 μM; NPEC1-Bmi1, 5 μM), U0126 (50 μM) and LY294002 (50 μM) for 30 min were infected with EBV for 2 h, and then cultured in the fresh medium for 48 h until FACS analysis for the infection efficiency. (f,g) Downregulation of EGFR and c-Met impaired EBV infection. HNE1 cells were transfected with siRNA duplexes targeting EGFR or c-Met for 48 h, followed by real-time PCR analysis for the expressions of EGFR and c-Met (f) or FACS analysis for EBV infection efficiency (g). (h) HRas partially rescued the inhibitory effect of Gefitinib on EBV infection. HNE1 cells transfected with pMSCV-HRAS-V12 plasmid for 24 h were pre-incubated with 20 μM Gefitinib for 30 min, followed by exposure to EBV. (i) HRas failed to rescue the suppressive effect of silencing NRP1 on EBV infection. After transfected with siNRP1 or siControl for 36 h, HNE1 cells were reseeded, and then transfected with the plasmid for HRAS-V12 or the empty vector for 24 h, followed by exposure to EBV. The infected cells were analysed by FACS, with controls set to 100%. For (e–i), data represent three times independent experiments. Graphs show mean±s.e.m. ***P<0.001; **P<0.01; *P<0.05; Student’s t-test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25670642), licensed under a CC-BY license. Not internally tested by R&D Systems.

Knockdown Validated Detection of Human Neuropilin-1 by Knockdown Validated View Larger

Detection of Human Neuropilin-1 by Knockdown Validated NRP1 enhances EBV infection, while NRP2 suppresses EBV infection.(a,b) Downregulation of NRP1 impaired, whereas knockdown of NRP2 promoted EBV infection. HNE1 cells were transfected with siRNA duplexes targeting NRP1 or NRP2 for 48 h, followed by NRP expression analysis by real-time PCR (a) or analysis for the efficiency of EBV infection (b); n=3. (c) EBV infection was blocked by soluble NRP1ABC, but enhanced by antibody against NRP2. For NRP1ABC protein-blocking experiment, HNE1 cells were infected with EBV, which was pre-incubated with purified NRP1ABC for 1 h. For antibody against NRP2-blocking experiment, HNE1 cells were pre-incubated with an anti-NRP2 antibody (100 μg ml−1) or goat IgG (control) at 4 °C for 1 h and then were exposed to EBV at an MOI of 5 × 103 for 3 h at 4 °C. (d,e) Overexpression of NRP1 enhanced EBV infection, while NRP2 suppressed EBV infection. HNE1 cells were transiently transfected with the expression plasmid for NRP1, NRP2 or the empty vector (pMSCV) for 24 h, followed by analysis for the expression of NRP1 and NRP2 by western blotting (d) or were exposed to EBV (e). (f,g) EGF upregulated NRP1 expression and enhanced EBV infection. HNE1 cells cultured with 10 ng ml−1 EGF for 24 h were analysed for the expression of NRP1 and NRP2 by western blotting (f) or were exposed to EBV (g). (h,i) EGF-enhanced EBV infection was partially dependent on NRP1. After transfected with siRNA against NRP1 for 48 h, EGF-treated HNE1 and NPEC-Bmi1 cells maintained in KSF medium supplemented with EGF were analysed for NRP1 expression by western blotting (h) or were exposed to EBV (i). For (b), (c), (e), (g) and (i), HNE1 or NPEC1-Bmi1 cells were exposed to EBV at an MOI of 2.5 × 103 for 2 h at 37 °C, unless otherwise indicated. The percentage of GFP-positive infected cells was analysed by FACS 48 h post infection, with controls (empty vector-transfected cells or vehicle treated-cells) set to 100%. Data represent three to five independent experiments. Values in all graphs are means±s.e.m. ***P<0.001; **P<0.01; *P<0.05; Student’s t-test. For (d), (f) and (h), GAPDH served as an internal control. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25670642), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

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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); 100286859 (Porcine)
Alternate Names
BDCA4; BDCA-4; CD304 antigen; CD304; DKFZp686A03134; DKFZp781F1414; neuropilin 1; Neuropilin1; Neuropilin-1; NP1; NRP; NRP1; transmembrane receptor; Vascular endothelial cell growth factor 165 receptor; VEGF165R

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Product Specific Notices

This product or the use of this product is covered by U.S. Patents owned by The Regents of the University of California. This product is for research use only and is not to be used for commercial purposes. Use of this product to produce products for sale or for diagnostic, therapeutic or drug discovery purposes is prohibited. In order to obtain a license to use this product for such purposes, contact The Regents of the University of California.

U.S. Patent # 6,054,293, 6,623,738, and other U.S. and international patents pending.

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.

13 Citations: Showing 1 - 10
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  1. alpha2,6 Sialylation mediated by ST6GAL1 promotes glioblastoma growth
    Authors: S Gc, K Tuy, L Rickenback, R Jones, A Chakrabort, CR Miller, EA Beierle, VS Hanumanthu, AN Tran, JA Mobley, SL Bellis, AB Hjelmeland
    JCI Insight, 2022-11-08;7(21):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. Neuropilin 1 and its inhibitory ligand mini-tryptophanyl-tRNA synthetase inversely regulate VE-cadherin turnover and vascular permeability
    Authors: N Gioelli, LJ Neilson, N Wei, G Villari, W Chen, B Kuhle, M Ehling, F Maione, S Willox, S Brundu, D Avanzato, G Koulouras, M Mazzone, E Giraudo, XL Yang, D Valdembri, S Zanivan, G Serini
    Nature Communications, 2022-07-20;13(1):4188.
    Species: Human
    Sample Types: Whole Cells
    Applications: ELISA Capture
  3. Differential plasmacytoid dendritic cell phenotype and type I Interferon response in asymptomatic and severe COVID-19 infection
    Authors: M Severa, RA Diotti, MP Etna, F Rizzo, S Fiore, D Ricci, M Iannetta, A Sinigaglia, A Lodi, N Mancini, E Criscuolo, M Clementi, M Andreoni, S Balducci, L Barzon, P Stefanelli, N Clementi, EM Coccia
    PloS Pathogens, 2021-09-02;17(9):e1009878.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  4. SARS-CoV-2 Infects Syncytiotrophoblast and Activates Inflammatory Responses in the Placenta
    Authors: LB Argueta, LA Lacko, Y Bram, T Tada, L Carrau, T Zhang, S Uhl, BC Lubor, V Chandar, C Gil, W Zhang, B Dodson, J Bastiaans, M Prabhu, CM Salvatore, YJ Yang, RN Baergen, BR tenOever, NR Landau, S Chen, RE Schwartz, H Stuhlmann
    bioRxiv : the preprint server for biology, 2021-06-17;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Functional Assay
  5. Semaphorin 3F signaling actively retains neutrophils at sites of inflammation
    Authors: T Plant, S Eamsamarng, MA Sanchez-Ga, L Reyes, SA Renshaw, P Coelho, AS Mirchandan, JM Morgan, FE Ellett, T Morrison, D Humphries, ER Watts, F Murphy, XL Raffo-Irao, A Zhang, JL Cash, C Loynes, PM Elks, F Van Eeden, LM Carlin, AJW Furley, MKB Whyte, SR Walmsley
    J. Clin. Invest., 2020-06-01;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  6. Atypical chemokine receptor ACKR3/CXCR7 controls postnatal vasculogenesis and arterial specification by mesenchymal stem cells via Notch signaling
    Authors: ST Wei, YC Huang, ML Hsieh, YJ Lin, WC Shyu, HC Chen, CH Hsieh
    Cell Death Dis, 2020-05-04;11(5):307.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Western Blot
  7. iRGD synergizes with PD-1 knockout immunotherapy by enhancing lymphocyte infiltration in gastric cancer
    Authors: N Ding, Z Zou, H Sha, S Su, H Qian, F Meng, F Chen, S Du, S Zhou, H Chen, L Zhang, J Yang, J Wei, B Liu
    Nat Commun, 2019-03-22;10(1):1336.
    Species: Human
    Sample Types: Spheroids
    Applications: Neutralization
  8. Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output
    Authors: SL Giandomeni, SB Mierau, GM Gibbons, LMD Wenger, L Masullo, T Sit, M Sutcliffe, J Boulanger, M Tripodi, E Derivery, O Paulsen, A Lakatos, MA Lancaster
    Nat. Neurosci., 2019-03-18;22(4):669-679.
    Species: Human
    Sample Types: Organoid
    Applications: IHC
  9. 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., 2017-05-03;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: ICC
  10. Conformational remodeling of the fibronectin matrix selectively regulates VEGF signaling.
    Authors: Ambesi A, McKeown-Longo P
    J Cell Sci, 2014-06-30;127(0):3805-16.
    Species: Human
    Sample Types: Whole Cells
    Applications: Western Blot
  11. 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, 2014-01-02;9(1):e84179.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC
  12. 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., 2011-05-01;435(3):609-18.
    Species: Human
    Sample Types: Cell Lysates, Whole Cells
    Applications: Flow Cytometry, Western Blot
  13. 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., 2009-10-16;284(49):33966-81.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot

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Reviews for Human Neuropilin-1 Antibody

Average Rating: 3.8 (Based on 5 Reviews)

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Human Neuropilin-1 Antibody
By Anonymous on 04/08/2022
Application: WB Sample Tested: HEK293T human embryonic kidney cell line Species: Human

Human Neuropilin-1 Antibody
By Karina Kinghorn on 07/16/2021
Application: Immunocytochemistry/Immunofluorescence Sample Tested: HUVEC human umbilical vein endothelial cells Species: Human

Human Neuropilin-1 Antibody
By Karina Kinghorn on 03/24/2021
Application: WB Sample Tested: HUVEC human umbilical vein endothelial cells Species: Human

Human Neuropilin-1 Antibody
By Anonymous on 12/18/2018
Application: WB Sample Tested: HEK293T human embryonic kidney cell line Species: Human

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