|Nogo‑B in Mouse Embryonic Retina. Nogo‑B was detected in immersion fixed frozen sections of mouse embryonic retina (E13.5) using Sheep Anti-Mouse Nogo‑B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6596) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to retina. View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.|
|Detection of Mouse Nogo‑B by Western Blot. Western blot shows lysates of Neuro‑2A mouse neuroblastoma cell line and C2C12 mouse myoblast cell line. PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Mouse Nogo‑B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6596) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Nogo‑B at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.|
Nogo-B (A No-Go for neurite outgrowth Isoform B; also reticulon-4 isoform 2 and RTN-xS) is a 49-51 kDa member of the reticulon protein family. It is widely expressed, being reported everywhere but liver. Nogo-B appears to be principally expressed in the ER, but does have a receptor (NgBR) on cell surfaces. Intracellularly, Nogo-B is known to interact with Bcl-xl and Bcl-2, and may play in role in both apoptosis and angiogenesis. Mouse NOGO-B/B1 is a two transmembrane, 356 amino acid protein. It contains an N-terminal cytoplasmic domain (aa 1-182), two transmembrane segments (aa 183-203 and 273-293), an intervening luminal region (aa 204-272) and a C-terminal cytoplasmic domain (aa 294-356). Nogo-B contains multiple phosphorylation sites and undergoes acetylation. Caspase-7 cleaves Nogo-B between Asp15Ser16, generating a 40-42 kDa fragment. Nogo-B exists in two forms; B1 represents aa 1-168 spliced to aa 975-1162 of Nogo-A, while B2 represents aa 1-187 spliced to aa 975-1162 of Nogo-A. Over aa 1-183, mouse Nogo-B/B1 shares 76% aa identity with human Nogo-B/B1.
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