Detects mouse Contactin-6 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant human (rh) Contactin‑6 is observed, and less than 5% cross-reactivity with recombinant mouse (rm) Contactin-3, rmContactin-4, rmContactin‑5, and rhTAG-1 is observed.
Polyclonal Sheep IgG
Chinese hamster ovary cell line CHO-derived recombinant mouse Contactin-6 Glu20-Ser998 Accession # Q9JMB8
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.
Detection of Mouse Contactin‑6 by Western Blot.
Western blot shows lysates of mouse brain (cortex) tissue, mouse brain (hypothalamus) tissue, and mouse brain (cerebellum) tissue. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Mouse Contactin‑6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5890) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Contactin‑6 at approximately 130 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Contactin‑6 in Mouse Spinal Cord.
Contactin‑6 was detected in perfusion fixed frozen sections of adult mouse spinal cord using Sheep Anti-Mouse Contactin‑6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5890) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to gray matter. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.
Detection of Mouse Contactin‑6 by Simple WesternTM.
Simple Western lane view shows lysates of mouse brain (cortex) tissue and mouse brain (cerebellum) tissue, loaded at 0.2 mg/mL. A specific band was detected for Contactin‑6 at approximately 168 kDa (as indicated) using 10 µg/mL of Sheep Anti-Mouse Contactin‑6 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5890) followed by 1:50 dilution of HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
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.
Contactin-6, also known as NB-3, is a 130 kDa member of the TAG/F3 subgroup of Contactin GPI-linked neuronal adhesion proteins (1, 2). Mature mouse Contactin-6 consists of six immunoglobulin-like domains, a flexible linker region, four fibronectin type III domains, and a GPI anchor (3-5). Mouse Contactin-6 shares 90% and 96% amino acid (aa) sequence identity with human and rat Contactin-6, respectively. Alternate splicing generates an isoform with a 17 aa deletion in the first Ig-like domain (3). Contactin-6 expression is upregulated postnatally in the cerebellum, brainstem, and hippocampus, and it shows differential expression within various structures of the brain (3, 5-7). In the cerebellum, it is presynaptically localized at synapses between excitatory glutamatergic parallel fibers and Purkinje cells (7). Contactin-6 associates in cis with CHL-1 and in trans with Notch-1 on oligodendrocytes (6, 8). Its binding to Notch-1 triggers translocation of the Notch intracellular domain to the nucleus and promotes oligodendrogenesis (6). Contactin-6 function is important in the postnatal development and function of the cerebellum, as shown in knockout mice which exhibit poor motor coordination (5, 7).
Shimoda, Y. and K. Watanabe (2009) Cell Adhesion Migration 3:64.
Hu, Q.D. et al. (2006) Dev. Neurosci. 28:25.
Lee, S. et al. (2000) Gene 245:253.
Ogawa, J. et al. (1996) Neurosci. Lett. 218:173.
Takeda, Y. et al. (2003) J. Neurobiol. 56:252.
Cui, X.-Y. et al. (2004) J. Biol. Chem. 279:25858.
Sakurai, K. et al. (2009) Dev. Neurobiol. Aug 11 epub.
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but also provides information about sample types, species, and experimental conditions.
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