Nogo-B Antibody [Unconjugated]

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 (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 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 (CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; 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.

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

Nogo-B Antibody [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Nogo-B
  Met1-Val200
  Accession # NP_722550
• Specificity: Detects human and mouse Nogo-B in direct ELISAs and Western blots.
• 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: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

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

Alternate Names for Nogo-B Antibody [Unconjugated]

• 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. 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.

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.

Publications for Nogo-B Antibody (AF6034)(6)

Publications using AF6034

• Rubinelli, L;Manzo, OL;Sungho, J;Del Gaudio, I;Bareja, R;Marino, A;Palikhe, S;Di Mauro, V;Bucci, M;Falcone, DJ;Elemento, O;Ersoy, B;Diano, S;Sasset, L;Di Lorenzo, A; Suppression of endothelial ceramide de novo biosynthesis by Nogo-B contributes to cardiometabolic diseases Nature communications 2025-02-25 [PMID: 40000621] (Immunohistochemistry, Western Blot, Mouse)
• Bowen Wang, Zhiheng Zhao, Michael Xiong, Rui Yan, Ke Xu The endoplasmic reticulum adopts two distinct tubule forms Proceedings of the National Academy of Sciences 5/3/2022 [PMID: 35471903]
• Bowen Wang, Zhiheng Zhao, Michael Xiong, Rui Yan, Ke Xu The endoplasmic reticulum adopts two distinct tubule forms Proceedings of the National Academy of Sciences 2022-05-03 [PMID: 35471903] (Immunocytochemistry, Primate - Chlorocebus aethiops (African Green Monkey))
• NM Sayles, N Southwell, K McAvoy, K Kim, A Pesini, CJ Anderson, C Quinzii, S Cloonan, H Kawamata, G Manfredi Mutant CHCHD10 causes an extensive metabolic rewiring that precedes OXPHOS dysfunction in a murine model of mitochondrial cardiomyopathy Cell Reports, 2022-03-08;38(10):110475. 2022-03-08 [PMID: 35263592]
• Ying Zhu, Meng Yang, Xue-Hui Li, Wu-Jian Xu, Wei Gao, Yu-Han Chen et al. Nogo-B promotes epithelial-mesenchymal transition in lung fibrosis via PERK branch of the endoplasmic reticulum stress pathway Annals of Translational Medicine 5/14/2021 [PMID: 33987261]
• Ying Zhu, Meng Yang, Xue-Hui Li, Wu-Jian Xu, Wei Gao, Yu-Han Chen et al. Nogo-B promotes epithelial-mesenchymal transition in lung fibrosis via PERK branch of the endoplasmic reticulum stress pathway Annals of Translational Medicine 2021-05-14 [PMID: 33987261] (Immunohistochemistry, Western Blot, Mouse)

Bioinformatics

• Gene Symbol: RTN4
• Uniprot:
  • NP_722550()