Neuropilin-2 Antibody

R&D Systems | Catalog # AF567

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse, Rat

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Blockade of Receptor-ligand Interaction, Simple Western

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunocytochemistry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Neuropilin-2 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant rat Neuropilin‑2
Gln23-Asp857 (Val809-Asp825 del)
Accession # O35276

Specificity

Detects mouse and rat Neuropilin-2 in direct ELISAs and Western blots. In direct ELISAs, less than 10% cross‑reactivity with recombinant human Neuropilin-2 and recombinant rat Neuropilin-1 is observed.

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 Neuropilin-2 Antibody

Detection of Mouse and Rat Neuropilin-2 antibody by Western Blot.

Detection of Mouse and Rat Neuropilin‑2 by Western Blot.

Western blot shows lysates of C6 rat glioma cell line, LL/2 mouse Lewis lung carcinoma cell line, and bEnd.3 mouse endothelioma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Mouse/Rat Neuropilin-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF567) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Neuropilin-2 at approximately 110 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Neuropilin-2 antibody in Rat Brain by Immunohistochemistry (IHC-Fr).

Neuropilin‑2 in Rat Brain.

Neuropilin-2 was detected in perfusion fixed frozen sections of rat brain using Goat Anti-Mouse/Rat Neuropilin-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF567) at 15 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of Mouse and Rat Neuropilin-2 antibody by Simple WesternTM.

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

Simple Western lane view shows lysates of 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 5 µg/mL of Goat Anti-Mouse/Rat Neuropilin-2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF567) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Detection of Neuropilin-2 by Western Blot

Detection of Neuropilin-2 by Western Blot

Neuropilins are expressed in the CP, the cortical plate, and other brain-surrounding tissues and released in the CSF.(A) In situ hybridization and immunolabeling of Nrp1 and Nrp2 on embryonic brain sections at E13.5. Expression of the class 3 Sema receptors was detected in the nascent CP (black arrowheads), the cortical plate, and meninges. No signal was detected in the cortical ventricular zone (asterisks). Higher magnifications of the cortex and CP colabeled with anti-Pecam highlight Nrp1/2 expression in endothelial cells of the CP and the perineural vascular plexus. The white arrowheads point to the Nrp1/Nrp2 protein signal at the apical border of epithelial cells in the developing CP. High magnifications of the choroid plexi are shown right. (B and C) Single-cell transcriptome analysis of embryonic cortical cells shows that cortical progenitor cells express neither Nrp1 (B) nor Nrp2 (C) between E12 and E15. Pseudotime describes the differentiation from an apical progenitor (AP) cell to a postmitotic neuron (N). (D and E) Immunoblotting on embryonic CSF using antibodies against Nrp1 (D) and Nrp2 (E) indicates that soluble forms of both receptors indicated with arrows are released into the brain fluid. Recombinant Nrp1 and Nrp2 ectodomains fused to Fc fragment were used as positive controls of immunodetection. Scale bars, 500 μm (A, left), 50 μm (A, middle), and 10 μm (A, right). CP, cortical plate; MZ, marginal zone; VZ, ventricular zone; M, meninges; PV, perineural vascular plexus. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36399562), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Neuropilin-2 Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 2.5-15 µg/mL of this antibody will block 50% of the binding of 10 ng/mL of Recombinant Human VEGF165 (Catalog # 293-VE) to immobilized Recombinant Rat Neuropilin-2 Fc Chimera (Catalog # 567-N2) coated at 5 µg/mL (100 µL/well). At 60 μg/mL, this antibody will block >90% of the binding.

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of rat brain

Simple Western

5 µg/mL
Sample: 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: C6 rat glioma cell line, LL/2 mouse Lewis lung carcinoma cell line, and bEnd.3 mouse endothelioma cell line

Reviewed Applications

Read 7 reviews rated 4.4 using AF567 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.

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-1 (Npn-1, previously known as Neuropilin) and Npn-2 (previously known as Npn-1-related molecule) are type I transmembrane proteins that bind members of the class III secreted semaphorin subfamily which are implicated in repulsive axon guidance. The extracellular domain of these proteins is composed of two N-terminal CUB (complement-binding) domains (domains a1 and a2), two domains with homology to coagulation factors V and VIII (domains b1 and b2) and a MAM (meprin) domain (domain c). In the absence of ligands, neuropilins can form homo- and hetero-oligomers via homophilic interactions of their MAM domains. At the amino acid sequence level, Npn-1and Npn-2 share 44% identity. Npn-1 and Npn-2 show different binding specificities for different members of the semaphorin family. The expression patterns of Npn-1 and Npn-2 in developing neurons of the central and peripheral nervous systems are largely, though not completely nonoverlapping. Npn‑1 and Npn-2 are also expressed by endothelial and tumor cells and have been shown to be isoform-specific receptors for VEGF165. Npn‑1 was also reported to bind PlGF-2 and the VEGF-like protein from of virus NZ2.

References

  1. Fujisawa, H. and T. Kitsukawa (1998) Curr. Opin. Neurobiol. 8:587.
  2. Neufeld, G. et al. (1999) FASEB J. 13:9.
  3. Poltorak, Z. et al. (2000) J. Biol. Chem. 275:18040.

Alternate Names

Neuropilin2

Entrez Gene IDs

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

Gene Symbol

NRP2

UniProt

O35276

Additional Neuropilin-2 Products

Product Documents for 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 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 Neuropilin-2 Antibody

Customer Reviews for Neuropilin-2 Antibody (7)

4.4 out of 5
7 Customer Ratings
5 Stars
57%
4 Stars
29%
3 Stars
14%
2 Stars
0%
1 Stars
0%

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Customer Images

Showing  1 - 5 of 7 reviews Showing All
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  • Mouse/Rat Neuropilin-2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: E12.5 mouse embryo fixed in 4% PFA
    Species: Mouse
    Verified Customer | Posted 12/03/2020
    Antibody was stained on E12.5 mouse sections (attached picture) as well as E9.5 mouse sections. Worked well. Note the positive stain in the veins and the negative signal in the adjacent arteries. Dilution used - 1:20
    Neuropilin-2 Antibody AF567
  • Mouse/Rat Neuropilin-2 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Embryonic tissue
    Species: Mouse
    Verified Customer | Posted 11/08/2016
    Neuropilin-2 Antibody AF567
  • Mouse/Rat Neuropilin-2 Antibody
    Name: Anonymous
    Application: Whole mount immunofluorescence
    Sample Tested: Whole mount eye tissue
    Species: Mouse
    Verified Customer | Posted 09/07/2016
    Whole mouse eye tissue was fixed in 2% PFA, blocked and incubated O/N in AF567 diluted 1:250. Signal was detected using an alexafluor 488-labeled donkey anti goat secondary
    Neuropilin-2 Antibody AF567
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Hippocampal neurons
    Species: Mouse
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Immunofluorescence
    Sample Tested: Hippocampal neurons and astrocytes
    Species: Mouse
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: RN22 cells
    Species: Rat
    Verified Customer | Posted 12/19/2014
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Primary hippocampal neurons
    Species: Mouse
    Verified Customer | Posted 12/19/2014

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Protocols

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FAQs

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Associated Pathways

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