Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Avian - Chicken, Primate - Chlorocebus pygerythrus (Vervet Monkey), Transgenic Mouse

Applications

Validated:

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

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Immunocytochemistry, Apoptosis, Co-Immunoprecipitation, Efferocytosis Assay, Stimulation, Transcriptional Analysis

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Neogenin Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant mouse Neogenin
Ala42-Ile1033 (Asp442-Leu461 del)
Accession # NP_032710

Specificity

Detects human and mouse Neogenin in direct ELISAs and 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 Neogenin Antibody

Detection of Human and Mouse Neogenin antibody by Western Blot.

Detection of Human and Mouse Neogenin by Western Blot.

Western blot shows lysates of 3T3-L1 mouse embryonic fibroblast adipose-like cell line, Jurkat human acute T cell leukemia cell line, and Hepa 1-6 mouse hepatoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human/Mouse Neogenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1079) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Neogenin at approximately 230 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Neogenin antibody in Mouse Embryo by Immunohistochemistry (IHC-Fr).

Neogenin in Mouse Embryo.

Neogenin was detected in perfusion fixed frozen sections of mouse embryo (15 d.p.c.) using Goat Anti-Human/Mouse Neogenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1079) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to developing neurons. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Detection of Mouse Neogenin antibody by Simple WesternTM.

Detection of Mouse Neogenin by Simple WesternTM.

Simple Western lane view shows lysates of beta TC-6 mouse beta cell insulinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Neogenin at approximately 207 kDa (as indicated) using 10 µg/mL of Goat Anti-Human/Mouse Neogenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1079) 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 Neogenin by Immunohistochemistry

Detection of Neogenin by Immunohistochemistry

Subregion-specific expression of RGMs and Neogenin in the hippocampus.In situ hybridization (A–C, J–L) and immunohistochemistry (D–I, M–O) on coronal mouse brain sections at E16.5 (A–I) and P5 (J–O). Sections in D–I and M–O are counterstained in blue with fluorescent Nissl. (A–F) RGMa mRNA and protein are expressed in the ventricular zone (VZ), dentate gyrus (DG) and cornu ammonis (CA) region. Strong expression of RGMb mRNA and protein is detected in the pial surface lining the hippocampal fissure (HF). Neogenin transcripts and protein are widely expressed in the developing hippocampus (Hip). (G–I) Immunostaining with isotype matched controls. (J–L) In situ hybridization at P5 shows strong but differential expression patterns of RGMa, RGMb and Neogenin in the CA pyramidal cell layers (Pyr). In addition, strong expression of Neogenin is detected in the granular layer (GC) of the DG. (M–O) Immunohistochemistry reveals expression of RGMa and weak expression of RGMb in the stratum lacunosum moleculare (SLM) and fimbria (FIM). Neogenin strongly labels different hippocampal layers. CX, cortex; Hb, habenula; HC, hippocampal commissure; PO, polymorph layer; SO, stratum oriens; SR, stratum radiatum; Th, thalamus. Scale bar A–C: 400 µm, D–F: 300 µm, G–I: 300 µm, J–L: 500 µm and M–O: 400 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23457482), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Neogenin by Immunohistochemistry

Detection of Neogenin by Immunohistochemistry

RGM and Neogenin expression in the mouse olfactory system.In situ hybridization on coronal mouse brain sections at E16.5 (A–C′) and P5 (L–N′). Panels A′–C′ and L′–N′ show higher magnifications of boxed areas in A–C and L–N, respectively. Immunohistochemistry (D–F, H–J) and RGMa-AP section binding (G, K) on E16.5 coronal mouse brain sections. Sections in D–F and H–J are counterstained in blue with fluorescent Nissl. (A–C′) In situ hybridization shows differential expression patterns of RGMa, RGMb and Neogenin in the olfactory bulb and olfactory epithelium (OE). In line with this, immunohistochemistry reveals that axons of olfactory sensory neurons in the OE stain strongly for RGMb and weakly for RGMa and Neogenin. Furthermore, RGMa, RGMb and Neogenin are expressed on olfactory bulb axon projections such as the lateral olfactory tract (LOT). a, apical; ACa, anterior commissure pars anterior; AOB, accessory olfactory bulb; b, basal; CP, cortical plate; CRP, cribriform plate; EPL, external plexiform layer; GL, glomerular layer; GR, granule cell layer; IPL, internal plexiform layer; IZ, intermediate zone; LV, lateral ventricle; MCL, mitral cell layer; ONL, olfactory nerve layer; OVZ, olfactory ventricular zone; S, septum; STR, striatum; VN, vomeronasal nerve. Scale bar A–C 200 µm, A′–C′ 100 µm, D–F 300 µm, G 500 µm, H–J 400 µm, K 500 µm, L–N 400 µm and L′–N′ 200 µm. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23457482), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse Neogenin Antibody

Application
Recommended Usage

Blockade of Receptor-ligand Interaction

In a functional ELISA, 2.5-10 µg/mL of this antibody will block 50% of the binding of 200 ng/mL of Recombinant Chicken Netrin-1 (Catalog # 128‑N1) to immobilized Recombinant Mouse Neogenin Fc Chimera (Catalog # 1079‑NE) coated at 5 µg/mL (100 µL/well). At 300 μg/mL, this antibody will block >90% of the binding.

Immunohistochemistry

5-15 µg/mL
Sample:  Perfusion fixed frozen sections of mouse embryo (15 d.p.c.)

Simple Western

10 µg/mL
Sample: beta TC‑6 mouse beta cell insulinoma cell line

Western Blot

0.5 µg/mL
Sample: 3T3‑L1 mouse embryonic fibroblast adipose-like cell line, Jurkat human acute T cell leukemia cell line, and Hepa 1‑6 mouse hepatoma cell line

Reviewed Applications

Read 1 review rated 4 using AF1079 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: Neogenin

Neogenin (NEO) is a type I transmembrane protein that is crucial for axonal guidance and neuronal migration. It is also involved in regulating differentiation programs in many embryonic and adult tissues (1). Mouse NEO is widely expressed in adult tissues and is expressed throughout the mid to late stages of gestation, in both neuronal and non-neuronal tissues. It is a member of the immunoglobulin (Ig) superfamily and is closely related to deleted in colorectal cancer (DCC). Mouse NEO cDNA encodes a 1493 amino acid residue (aa) precursor with a putative 36 aa signal peptide, a 1100 aa extracellular domain with six Ig-like C2 type domains and three fibronectin type III domains, a 21 aa transmembrane domain, and a 345 aa cytoplasmic domain. At least five isoforms are produced in mice by alternative splicing. Mouse NEO shares 96%, 93%, and 86% aa sequence identity with rat, human, and chicken NEO, respectively. It also has 46% and 29% sequence homology with mouse DCC and C. elegans UNC40, a homolog of DCC. NEO and DCC, together with the UNC5 family of type I transmembrane proteins, are receptors for the netrin/UNC6 family of laminin-related bifunctional guidance molecules that both attract some axons and repel others (2, 3). In mouse, at least five netrins (netrin‑1,
‑3, -4, G1, and G2) have been identified (3-5). Mouse netrin-1 and netrin-3 have been shown to be ligands for mouse NEO.

References

  1. Keeling, S.L. et al. (1997) Oncogene 15:691.
  2. Hong, K. et al. (1999) Cell 97:927.
  3. Livesey, F.J. (1999) Cell Mol. Life Sci. 56:62.
  4. Nakashiba, T. et al. (2000) J. Neurosci. 20:6540.
  5. Nakashiba, T. et al. (2002) Mech. Dev. 111:47.

Alternate Names

NEO, NEO1, NGN

Entrez Gene IDs

4756 (Human); 18007 (Mouse)

Gene Symbol

NEO1

UniProt

NP_032710

Additional Neogenin Products

Product Documents for Human/Mouse Neogenin Antibody

Certificate of Analysis

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Product Specific Notices for Human/Mouse Neogenin 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 # 5,939,271, 6,277,585, and other U.S. and international patents pending.

For research use only

Citations for Human/Mouse Neogenin Antibody

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  • Human/Mouse Neogenin Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Brain tissue
    Species: Mouse
    Verified Customer | Posted 01/25/2019
    Dilution used for the western is 1:250
    Human/Mouse Neogenin Antibody AF1079

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