hh Human Nogo-A aa 566-748 Antibody AF3515: R&D Systems

Human Nogo-A aa 566-748 Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Nogo-A in direct ELISAs and Western blots. In direct ELISAs, approximately 5% cross-reactivity with recombinant rat Nogo-A (aa 543-725) and recombinant human Nogo-B is observed..
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Chinese hamster ovary cell line CHO-derived recombinant human Nogo-A
    Val566-Phe748
    Accession # Q9NQC3
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.1 µg/mL
    Recombinant Human Nogo-A Fc Chimera, aa 566-748 (Catalog # 3515-NG)
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Immunocytochemistry

Nogo‑A in Rat Cortical Stem Cells.
Nogo‑A was detected in immersion fixed 7 day differentiated rat cortical stem cells using 10 µg/mL Sheep Anti-Human Nogo‑A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3515) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    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.
Background: Nogo-A

Human Nogo-A (also reticulon-4) is a member of the reticulon family of transmembrane proteins. This family is characterized by the presence of a nonsignal sequence-containing N-terminus, a topologically conserved 200 amino acid (aa) C-terminus that contains two transmembrane domains with an ER-retention motif, and a punctate intracellular distribution within the ER that is reminescent of a reticulum (1-4). In human, Nogo exists in five isoforms (5-7). The full length human form (Nogo-A) is 1192 aa in length and contains a 1018 aa N-terminus, a 21 aa transmembrane segment, a 94 aa connecting “loop”, a second 21 aa transmembrane segment, and a 38 aa C-terminus. Three areas are of particular interest. One is a stretch of 66 aa within the 94 aa connecting loop. This segment is reported to bind to the GPI-linked Nogo receptor/p75 complex on axons and induce growth cone collapse (8-10). Two other areas in the N-terminus have also been discovered to have bioactivity (8, 11, 12). Based on rat, aa 57-184 in human (aa 59-172 in rat) should block fibroblast spreading, while aa 566-748 in human (aa 544-725 in rat) block neurite outgrowth and block fibroblast spreading (8, 12, 13). The exact topology of Nogo-A is unclear. The N- and C-termini may be extracellular with the “loop” region intracellular, or the situation could be reversed (13-15). Alternatively, the loop region and N-terminus may be on the same side of the membrane (3, 8). The four additional isoforms are shorter than Nogo-A (199 aa [Nogo-C], 373 aa [Nogo-B], 392 aa and 986 aa, respectively) (7). Although highly divergent, all contain the same C-terminal stretch, aa 1005-1192. Both Nogo-B and C are reported to complex with Nogo-A (16). Notably, Nogo-A is expressed in neurons, endothelial cells. oligodendrocytes, fibroblasts and myoblasts (12, 16-18). Human Nogo-A is 78% aa identical to mouse and rat Nogo-A overall, with 98% aa identity in the loop region and approximately 80% aa identity in the aa 566-748 segment.

  • References:
    1. Oertle, T. et al. (2003) FASEB J. 17:1238.
    2. GrandPre, T. et al. (2000) Nature 403:439.
    3. Yan, R. et al. (2006) Cell. Mol. Life Sci. 63:877.
    4. Teng, F.Y.H. et al. (2004) J. Neurochem. 89:801.
    5. Chen, M.S. et al. (2000) Nature 403:434.
    6. Morris, N.J. et al. (1999) Biochim. Biophys. Acta 1450:68.
    7. GenBank Accession # Q9NQC3.
    8. Oertle, T. et al. (2003) J. Neurosci. 23:5393.
    9. Fournier, A.E. et al. (2001) Nature 409:341.
    10. Wang, K.C. et al. (2002) Nature 420:74.
    11. Prinjha, R. et al. (2000) Nature 403:384.
    12. Dodd, D.A. et al. (2005) J. Biol. Chem. 280:12494.
    13. Li, M. et al. (2004) Eur. J. Biochem. 271:3512.
    14. Huber, A.B. and M.E. Schwab (2000) Biol. Chem. 381:407.
    15. Ng, C.E.L. and B.L. Tang (2002) J. Neurosci. Res. 67:559.
    16. Dodd, D.A. et al. (2005) J. Biol. Chem. 280:12494.
    17. Wang, X. et al. (2002) J. Neurosci. 22:5505.
    18. Acevedo, L. et al. (2004) Nat. Med. 10:382.
  • Long Name:
    Reticulon 4A
  • Entrez Gene IDs:
    57142 (Human); 68585 (Mouse); 83765 (Rat)
  • Alternate Names:
    NI220; NogoA; Nogo-A; RTN4; RTN4A
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Protein microarray analysis identifies cyclic nucleotide phosphodiesterase as an interactor of Nogo-A.
    Authors: Sumiyoshi K, Obayashi S, Tabunoki H
    Neuropathology, 2010;30(1):7-14.
    Species: Mouse
    Sample Type: Whole Tissue
    Application: IHC Paraffin-embedded
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