Nogo-B Antibody

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.

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.

• Reactivity: Mu
• Applications: WB, IHC
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Nogo-B Antibody Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Nogo-B
  Met1-Val183
  Accession # NP_918941
• Specificity: 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.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: Rtn4
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Nogo-B Antibody

• ASY;Nbla00271;Nbla10545;NI220/250;NOGO;NSP;NSP-CL;Reticulon-4;RTN4;RTN4-A;RTN4-B1;RTN4-B2;RTN4-C;RTN-X
• NogoB
• Nogo-B
• RTN4B

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Gene Symbol: Rtn4
• Uniprot:
  • NP_918941()