Human/Mouse Neogenin Antibody

AF1079 8 Citations Datasheet / COA / SDS

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AF1079
AF1079-SP

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Detection of Human and Mouse Neogenin by Western Blot.

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Citations (8)

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human/Mouse Neogenin Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human and mouse Neogenin in direct ELISAs and Western blots. In Western blots, less than 5% cross-reactivity with recombinant mouse DCC is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

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

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

Endotoxin Level

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

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.5 µg/mL

See below

Simple Western

10 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

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.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot

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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.

Immunohistochemistry

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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.

Simple Western

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Detection of Mouse Neogenin by Simple Western^TM. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (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.

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

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

Entrez Gene IDs

4756 (Human); 18007 (Mouse)

Alternate Names

HsT17534; IGDCC2DKFZp547B146; Immunoglobulin superfamily DCC subclass member 2; NEO; NEO1; neogenin (chicken) homolog 1; neogenin 1; neogenin homolog 1; Neogenin; NGN; NGNDKFZp547A066

Product Datasheets

Product Datasheet

COA

SDS

Related Research Areas

Citations for Human/Mouse Neogenin Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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Tbr1 instructs laminar patterning of retinal ganglion cell dendrites
Authors: J Liu, JDS Reggiani, MA Laboulaye, S Pandey, B Chen, JLR Rubenstein, A Krishnaswa, JR Sanes
Nat. Neurosci., 2018;0(0):.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC
Effect of Erythropoietin, Iron Deficiency and Iron Overload on Liver Matriptase-2 (TMPRSS6) Protein Content in Mice and Rats.
Authors: Frydlova J, Prikryl P, Truksa J, Falke L, Du X, Gurieva I, Vokurka M, Krijt J
PLoS ONE, 2015;11(2):e0148540.
Species: Mouse
Sample Types: Tissue Homogenates
Applications: WB
The axonal guidance receptor neogenin promotes acute inflammation.
Authors: Konig K, Gatidou D, Granja T, Meier J, Rosenberger P, Mirakaj V
PLoS ONE, 2012;7(3):e32145.
Species: Mouse
Sample Types: In Vivo
Applications: In vivo
Genetic analysis of DSCAM's role as a Netrin-1 receptor in vertebrates.
Authors: Palmesino E, Haddick PC, Tessier-Lavigne M, Kania A
J. Neurosci., 2012;32(2):411-6.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC Frozen
Netrin-1 overexpression protects kidney from ischemia reperfusion injury by suppressing apoptosis.
Authors: Wang W, Reeves WB, Pays L, Mehlen P, Ramesh G
Am. J. Pathol., 2009;175(3):1010-8.
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC Paraffin-embedded
Interaction of amyloid precursor protein with contactins and NgCAM in the retinotectal system.
Authors: Osterfield M, Egelund R, Young LM, Flanagan JG
Development, 2008;135(6):1189-99.
Species: Chicken
Sample Types: Tissue Homogenates
Applications: WB
Selective binding of RGMc/hemojuvelin, a key protein in systemic iron metabolism, to BMP-2 and neogenin.
Authors: Kuns-Hashimoto R, Kuninger D, Nili M, Rotwein P
Am. J. Physiol., Cell Physiol., 2008;294(4):C994-C1003.
Species: Primate - Chlorocebus pygerythrus (Vervet Monkey)
Sample Types: Whole Cells
Applications: Neut
Netrin-4 inhibits angiogenesis via binding to neogenin and recruitment of Unc5B.
Authors: Lejmi E, Leconte L, Pedron-Mazoyer S, Ropert S, Raoul W, Lavalette S, Bouras I, Feron JG, Maitre-Boube M, Assayag F, Feumi C, Alemany M, Jie TX, Merkulova T, Poupon MF, Ruchoux MM, Tobelem G, Sennlaub F, Plouet J
Proc. Natl. Acad. Sci. U.S.A., 2008;105(34):12491-6.
Species: Human
Sample Types: Cell Lysates
Applications: WB