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Human Neuropilin-1 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF3870

R&D Systems, part of Bio-Techne
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AF3870
AF3870-SP

Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Blockade of Receptor-ligand Interaction, CyTOF-ready, Flow Cytometry, Immunocytochemistry, Immunohistochemistry, Western Blot

Cited:

ELISA Capture, Flow Cytometry, Functional Assay, Immunocytochemistry, Immunohistochemistry, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Neuropilin-1
Phe22-Lys644
Accession # NP_001019799

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Human Neuropilin-1 Antibody

Detection of Human Neuropilin-1 antibody by 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.
Detection of Neuropilin-1 antibody in HUVEC Human Cells antibody by 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).
Neuropilin-1 antibody in PC-3 and HLDM-2 Human Cell Lines by Immunocytochemistry (ICC).

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.

Applications for Human Neuropilin-1 Antibody

Application
Recommended Usage

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.

Flow Cytometry

2.5 µg/106 cells
Sample: HUVEC human umbilical vein endothelial cells

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed PC-3 human prostate cancer cell line and HLDM-2 human Hodgkin's lymphoma cell line

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human ovarian and pancreatic cancer tissue

Western Blot

1 µg/mL
Sample: MDA‑MB‑231 human breast cancer cell line and human placenta tissue
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 5 reviews rated 3.8 using AF3870 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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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

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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, Neuropilin1, NRP1

Entrez Gene IDs

8829 (Human); 18186 (Mouse); 246331 (Rat); 100286859 (Porcine)

Gene Symbol

NRP1

UniProt

Additional Neuropilin-1 Products

Product Documents for Human Neuropilin-1 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

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.

For research use only

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