Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

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

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Blocking, ELISA Development

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Neuropilin-2 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Neuropilin‑2
Gln23-Tyr855
Accession # Q7LBX6

Specificity

Detects human, mouse, and rat Neuropilin-2 in Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

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

Scientific Data Images for Human/Mouse/Rat Neuropilin‑2 Antibody

Detection of Human Neuropilin-2 antibody by Western Blot.

Detection of Human Neuropilin‑2 by Western Blot.

Western blot shows lysates of HUVEC human umbilical vein endothelial cells. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse/Rat Neuropilin-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2215) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody HAF109). A specific band was detected for Neuropilin-2 at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Neuropilin-2 antibody in Human Pancreatic Cancer Tissue by Immunohistochemistry (IHC-P).

Neuropilin‑2 in Human Pancreatic Cancer Tissue.

Neuropilin-2 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using Goat Anti-Human/Mouse/Rat Neuropilin-2 Antigen Affinity-purified Poly-clonal Antibody (Catalog # AF2215) at 5 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 (CTS013). Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human, Mouse, and Rat Neuropilin-2 antibody by Simple WesternTM.

Detection of Human, Mouse, and Rat Neuropilin‑2 by Simple WesternTM.

Simple Western lane view shows lysates of HUVEC human umbilical vein endothelial cells, C6 rat glioma cell line, LL/2 mouse Lewis lung carcinoma cell line, and bEnd.3 mouse endothelioma cell line, loaded at 0.2 mg/mL. A specific band was detected for Neuropilin-2 at approximately 140 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse/Rat Neuropilin-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2215) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Neuropillin-2 in HUVEC Human Cell Line by Flow Cytometry.

HUVEC human umbilical vein endothelial cells were stained with Goat Anti-Human/Mouse/Rat Neuropillin-2 Polyclonal Antibody (Catalog # AF2215, filled histogram) or Goat IgG control antibody (AB-108-C, open histogram), followed by Phycoerythrin-conjugated anti-Goat IgG (F0107). Staining was performed using our Staining Surface Molecules protocol.

Applications for Human/Mouse/Rat Neuropilin‑2 Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 1-10 µg/mL of this antibody will block 50% of the binding of 500 ng/mL of Recombinant Human VEGF165  (Catalog # 293-VE) to immobilized Recombinant Human Neuropilin-2 Fc Chimera  (Catalog # 2215-N2) coated at 1 µg/mL (100 µL/well). At 20 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

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

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human pancreatic cancer tissue subjected to Antigen Retrieval Reagent-Basic (Catalog # CTS013)

Simple Western

10 µg/mL
Sample: HUVEC human umbilical vein endothelial cells, C6 rat glioma cell line, LL/2 mouse Lewis lung carcinoma cell line, and bEnd.3 mouse endothelioma cell line

Western Blot

0.5 µg/mL
Sample: HUVEC human umbilical vein endothelial cells

Reviewed Applications

Read 2 reviews rated 3 using AF2215 in the following applications:

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

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.

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.

Calculators

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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Background: Neuropilin-2

Neuropilin-2 (Npn-2) is a 120 kDa, type I transmembrane (TM) glycoprotein that is related to the semaphorin receptor now known as Neuropilin-1 (1). Npn-2 is a complex molecule with multiple splice forms. Five transmembrane forms are known, and one 62 kDa soluble form has been identified (2). Based on the originally reported precursor size of 909 amino acids (aa), the “standard” precursor in human will have a 20 aa signal sequence, an 842 aa extracellular region, a 25 aa TM segment, and a 42 aa cytoplasmic tail (1). The extracellular region contains two N-terminal CUB (C1r/Ugef/BMP-1) domains, two jellyroll-shaped coagulation factor V type C domains, and a juxtamembrane MAM (meprin/A-5 protein/tyrosine phosphatase μ) domain (1, 3). The CUB and factor V domain are involved in VEGF and semaphorin binding. The MAM domain appears necessary for signaling through plexin-1 (4). The five transmembrane isoforms all share the same CUB, factor V and MAM domains. Splicing begins at aa 809, seven amino acids after the end of the MAM domain, and it involves the end of the extracellular region, the TM segment, and the cytoplasmic domain (a total of 101 aa). Two of the four variants show a complete replacement of these 101 aa with a totally unrelated stretch of approximately 90 aa. This creates a new TM and cytoplasmic tail. These forms are called “Npn-2b” forms. Two other isoforms (plus the standard 909 aa form) retain the 101 aa stretch, and add either 17 or 22 aa to the end of the extracellular region. These forms are called “Npn-2a” forms. The isoform offered by R&D Systems is the “a” form with the 17 aa addition. This isoform shows 94% aa identity to the equivalent regions in mouse and rat Npn-2. The soluble form of Npn-2 is 555 aa in precursor length, and contains the two CUB domains plus the first 1½ factor V type C domains (1). Npn-2 binds Sema3B through F, and VEGF isoforms 165, 145, PlGF-2, and VEGF-C (5). It is known to form homodimers and heterodimers with Npn-1, and it forms receptor complexes with plexin-1 and VEGF R1 (4, 5). Npn-2 is found on a variety of cell types including neurons (motor, autonomic, sensory), vascular endothelial cells, Schwann cells and pancreatic acinar cells.

References

  1. Chen, H. et al. (1997) Neuron 19:547.
  2. Rossignol, M. et al. (2000) Genomics 70:211.
  3. He, Z. and M. Tessier-lavigne (1997) Cell 90:739.
  4. Nakamura, F. and Y. Goshima (2002) Adv. Exp. Med. Biol. 515:55.
  5. Neufeld, G. et al. (2002) Adv. Exp. Med. Biol. 515:81.

Alternate Names

Neuropilin2

Entrez Gene IDs

8828 (Human); 18187 (Mouse); 81527 (Rat)

Gene Symbol

NRP2

UniProt

Q7LBX6

Additional Neuropilin-2 Products

Product Documents for Human/Mouse/Rat Neuropilin‑2 Antibody

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat Neuropilin‑2 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

Citations for Human/Mouse/Rat Neuropilin‑2 Antibody

Customer Reviews for Human/Mouse/Rat Neuropilin‑2 Antibody (2)

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2 Customer Ratings
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  • Name: Zhipeng Meng
    Verified Customer | Posted 04/21/2017
    Human/Mouse/Rat Neuropilin‑2 Antibody AF2215
  • Human/Mouse/Rat Neuropilin-2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Neonatal dorsal root ganglia cells
    Species: Mouse
    Verified Customer | Posted 09/01/2016

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Protocols

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Associated Pathways

VEGF - VEGF R2 Signaling Pathways VEGF - VEGF R2 Signaling Pathway Thumbnail