Detection of Human and Mouse Nogo‑B by Western Blot.
Western blot shows lysates of TK‑6 human lymphoblast cell line, Neuro‑2A mouse neuroblastoma cell line, and C2C12 mouse myoblast cell line. PVDF Membrane was probed with 1 µg/mL of Human/Mouse Nogo‑B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6034) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Nogo‑B at approximately 49 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
Nogo-B in Human Heart.
Nogo-B was detected in immersion fixed paraffin-embedded sections of human heart using Human/Mouse Nogo-B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6034) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). 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 endothelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
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. Human Nogo-B is a two transmembrane, 373 amino acid protein. It contains an N-terminal cytoplasmic domain (aa 1‑199), two transmembrane segments (aa 200‑219 and 315‑335), a luminal region (aa 220‑314) and a C-terminal cytoplasmic domain (aa 336‑373). Nogo-B contains multiple phosphorylation sites and undergoes acetylation. Caspase-7 cleaves Nogo‑B between Asp14Ser15, generating a 45‑47 kDa fragment. Nogo-B exists in two forms; B1 represents aa 1‑185 spliced to aa 1005‑1192 of Nogo-A, while B2 represents aa 1‑204 spliced to aa 1005‑1192 of Nogo-A. Over aa 1‑200 (of Nogo-A), human Nogo-B shares 76% aa identity with mouse Nogo-B.
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