Human/Mouse Neurexin 3/NRXN3 Antibody

Catalog #: AF5269
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AF5269
AF5269-SP

Detection of Neurexin 3/NRXN3 in Human Brain Cerebellum.

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Human/Mouse Neurexin 3/NRXN3 Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human and mouse Neurexin 3/NRXN3 in Western blots.

Source

Polyclonal Sheep IgG

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

2 µg/mL

Mouse brain

Immunohistochemistry

5-15 µg/mL

Immersion fixed paraffin-embedded sections of human brain (cerebellum and cortex)

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.

Scientific Data

Immunohistochemistry

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Detection of Neurexin 3/NRXN3 in Human Brain Cerebellum. Neurexin 3/NRXN3 was detected in immersion fixed paraffin-embedded sections of human brain cerebellum using Sheep Anti-Human/Mouse Neurexin 3/NRXN3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5269) at 15 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Sheep IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC006). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to Purkinje Neurons. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Western Blot

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Detection of Human/Mouse Neurexin 3/NRXN3 by Western Blot. Western blot shows lysates of Mouse Brain. PVDF membrane was probed with 2 µg/mL of Sheep Anti-Human/Mouse Neurexin 3/NRXN3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5269) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Neurexin 3/NRXN3 at approximately ~150 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

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

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

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Neurexin 3/NRXN3

The alpha and beta forms of Neurexins 1‑3 are transmembrane neuronal glycoproteins which are transcribed from each of three NRXN genes that utilize alternate promoters. Like other Neurexins, the extracellular domain (ECD) of Neurexin 3 alpha contains six LNS domains interspersed with three EGF-like domains, while that of Neurexin 3 beta contains only the sixth LNS domain and no EGF-like domains (1‑3). Mature human Neurexin 3 beta is a 70 kDa glycosylated protein with a 528 amino acid (aa) ECD and a 56 aa cytoplasmic domain that contains a motif for binding PDZ scaffolding proteins (3‑5). Within comparable regions of the ECD, human Neurexin 3 beta shares 99% aa sequence identity with mouse and rat Neurexin 3 beta. It shares 65% aa sequence identity with comparable regions of the ECD of human Neurexin 1 beta and 2 beta. Alternative splicing of human Neurexin 3 beta generates multiple isoforms. There are potentially soluble and secreted variants and some which contain a fibronectin type III-like domain (4, 6). Neurexin 3 beta is widely expressed in the brain where it binds the postsynaptic Neuroligins 1, 2, and 3 (6‑9). Neurexin 3 beta may also be expressed in non-nervous tissues with a potentially cardiac-specific isoform (10). Human Neurexin 3 beta polymorphisms which affect the splicing pattern are associated with susceptibility to alcohol dependence (6). The Neurexin 3 beta genetic locus has been linked to opioid and nicotine addiction, and Neurexin 3 beta gene expression is up‑regulated after short term exposure of mice to cocaine (11‑13).

References

Craig, A.M. and Y. Kang (2007) Curr. Opin. Neurobiol. 17:43.
Dean, C. and T. Dresbach (2006) Trends Neurosci. 29:21.
Lise, M.-F. and A. El-Husseini (2006) Cell. Mol. Life Sci. 63:1833.
Rowen, L. et al. (2002) Genomics 79:587.
Ushkaryov, Y. A. et al. (1994) J. Biol. Chem. 269:11987.
Hishimoto, A. et al. (2007) Hum. Mol. Genet. 16:2880.
Ullrich, B. et al. (1995) Neuron 14:497.
Ichtchenko, K. et al. (1996) J. Biol. Chem. 271:2676.
Ichtchenko, K. et al. (1995) Cell 81:435.
Occhi, G. et al. (2002) Biochem. Biophys. Res. Commun. 298:151.
Lachman, H.M. et al. (2007) Hum. Mol. Genet. 16:1327.
Bierut, L.J. et al. (2007) Hum. Mol. Genet. 16:24.
Kelai, S. et al. (2008) Neuroreport 19:751.

Entrez Gene IDs

9369 (Human); 18191 (Mouse)

Alternate Names

C14orf60; chromosome 14 open reading frame 60; KIAA0743MGC176711; Neurexin 3; neurexin III; Neurexin III-alpha; Neurexin III-beta; neurexin-3-beta; NRXN3

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Citation for Human/Mouse Neurexin 3/NRXN3 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.

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