Human Neuropilin-1 Antibody

Name: Human Neuropilin-1 Antibody
Brand: R&D Systems
SKU: AF3870
Rating: 4 (5 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AF3870

Citations (13)

(5)

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Key Product Details

Species Reactivity:	Human
Immunogen:	Mouse myeloma cell line NS0-derived recombinant human Neuropilin‑1 Phe22-Lys644 Accession # NP_001019799
Applications:	Block/Neutralize, CyTOF-ready, Flow Cytometry, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Western Blot
Label:	Unconjugated
Source:	Polyclonal Sheep IgG
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.

View Full Technical Details for Human Neuropilin-1 Antibody

Applications for Human Neuropilin-1 Antibody

	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.

Scientific Data Examples for Human Neuropilin-1 Antibody

	Western Blot 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. 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 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 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.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Endotoxin level	<0.10 EU per 1 μg of the antibody by the LAL method.
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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.
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.

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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).

Item	Value
References	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. Yaron, A. et al. (2005) Neuron 45:513. Narazaki, M. and G. Tosato (2006) Blood 107:3892. Tordjman, R. et al. (2002) Nat. Immunol. 3477.
Entrez Gene IDs	8829 (Human); 18186 (Mouse); 246331 (Rat); 100286859 (Porcine)
Alternate Names	BDCA-4, BDCA4, CD304, CD304 antigen, DKFZp686A03134, DKFZp781F1414, NP1, NRP, NRP1, Neuropilin-1, Neuropilin1, VEGF165R, Vascular endothelial cell growth factor 165 receptor, neuropilin 1, transmembrane receptor

Product Specific Notices for Human Neuropilin-1 Antibody

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 (13)

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Citations are publications that use Bio-Techne products. Selected citations for Human Neuropilin-1 Antibody include:

Species: Human
Sample Types: Whole Cells
Applications: ELISA Capture

N Gioelli et al. (2022), Neuropilin 1 and its inhibitory ligand mini-tryptophanyl-tRNA synthetase inversely regulate VE-cadherin turnover and vascular permeability Nature Communications, 13(1):4188.
PMID: 35858913
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

S Gc et al. (2022), alpha2,6 Sialylation mediated by ST6GAL1 promotes glioblastoma growth JCI Insight, 7(21).
PMID: 36345944
Species: Human
Sample Types: Whole Cells
Applications: Functional Assay

LB Argueta et al. (2021), SARS-CoV-2 Infects Syncytiotrophoblast and Activates Inflammatory Responses in the Placenta bioRxiv : the preprint server for biology, 0(0).
PMID: 34100019
Species: Human
Sample Types: Whole Cells
Applications: Neutralization

M Severa et al. (2021), Differential plasmacytoid dendritic cell phenotype and type I Interferon response in asymptomatic and severe COVID-19 infection PloS Pathogens, 17(9):e1009878.
PMID: 34473805
Species: Human
Sample Types: Cell Culture Supernates
Applications: Western Blot

ST Wei et al. (2020), Atypical chemokine receptor ACKR3/CXCR7 controls postnatal vasculogenesis and arterial specification by mesenchymal stem cells via Notch signaling Cell Death Dis, 11(5):307.
PMID: 32366833
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC

T Plant et al. (2020), Semaphorin 3F signaling actively retains neutrophils at sites of inflammation J. Clin. Invest., 0(0).
PMID: 32191647
Species: Human
Sample Types: Spheroids
Applications: Neutralization

N Ding et al. (2019), iRGD synergizes with PD-1 knockout immunotherapy by enhancing lymphocyte infiltration in gastric cancer Nat Commun, 10(1):1336.
PMID: 30902997
Species: Human
Sample Types: Organoid
Applications: IHC

SL Giandomeni et al. (2019), Cerebral organoids at the air-liquid interface generate diverse nerve tracts with functional output Nat. Neurosci., 22(4):669-679.
PMID: 30886407
Species: Human
Sample Types: Whole Cells
Applications: ICC

C Kerros et al. (2017), Neuropilin-1 mediates Neutrophil Elastase uptake and cross-presentation in breast cancer cells J. Biol. Chem., 0(0).
PMID: 28468826
Species: Human
Sample Types: Whole Cells
Applications: Western Blot

Ambesi A et al. (2014), Conformational remodeling of the fibronectin matrix selectively regulates VEGF signaling. J Cell Sci, 127(0):3805-16.
PMID: 24982443
Species: Mouse
Sample Types: Whole Cells
Applications: ICC

Boyer-Di Ponio J et al. (2014), Instruction of circulating endothelial progenitors in vitro towards specialized blood-brain barrier and arterial phenotypes. PLoS ONE, 9(1):e84179.
PMID: 24392113
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Pellet-Many C et al. (2011), Neuropilin-1 mediates PDGF stimulation of vascular smooth muscle cell migration and signalling via p130Cas. Biochem. J., 435(3):609-18.
PMID: 21306301
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot

Robinson SD et al. (2009), Alphav beta3 integrin limits the contribution of neuropilin-1 to vascular endothelial growth factor-induced angiogenesis. J. Biol. Chem., 284(49):33966-81.
PMID: 19837659

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Human Neuropilin-1 Antibody

Name: Anonymous

Application: WB

Sample Tested: HEK293T human embryonic kidney cell line

Species: Human

Verified Customer | Posted 04/08/2022
Human Neuropilin-1 Antibody

Name: Karina Kinghorn

Application: ICC/IF

Sample Tested: HUVEC human umbilical vein endothelial cells

Species: Human

Verified Customer | Posted 07/16/2021
Human Neuropilin-1 Antibody

Name: Karina Kinghorn

Application: WB

Sample Tested: HUVEC human umbilical vein endothelial cells

Species: Human

Verified Customer | Posted 03/24/2021
Human Neuropilin-1 Antibody

Name: Anonymous

Application: WB

Sample Tested: HEK293T human embryonic kidney cell line

Species: Human

Verified Customer | Posted 12/18/2018
Name: Anonymous

Verified Customer | Posted 04/21/2017

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Human Neuropilin-1 Antibody

R&D Systems, Inc. a Bio-Techne Brand

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Applications for Human Neuropilin-1 Antibody

Scientific Data Examples for Human Neuropilin-1 Antibody

Western Blot

Flow Cytometry

Immunocytochemistry

Immunohistochemistry

Formulation, Preparation and Storage

Reconstitution Calculator

Background: Neuropilin-1

Product Specific Notices for Human Neuropilin-1 Antibody

Citations for Human Neuropilin-1 Antibody (13)

Customer Reviews for Human Neuropilin-1 Antibody (5)

Customer Images

Human Neuropilin-1 Antibody

Human Neuropilin-1 Antibody

Human Neuropilin-1 Antibody

Human Neuropilin-1 Antibody

Product Documents for Human Neuropilin-1 Antibody

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