Detects mouse Nogo-B in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant human (rh) Nogo-B and recombinant rat (rr) Nogo-A (aa 2-172) is observed and less than 1% cross-reactivity with rhNogo-A, rrNogo-A (aa 544-725), and rrNogo-A (aa 1026-1091) is observed.
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.
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.
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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