Nogo-A Antibody

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.

• Reactivity: Hu
• Applications: WB, ICC/IF
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Nogo-A Antibody Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Nogo-A
  Val566-Phe748
  Accession # Q9NQC3
• 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: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: RTN4
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry 5-15 ug/mL
  • Western Blot 0.1 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 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC 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-A Antibody

• ASY;Nbla00271;Nbla10545;NI220/250;Nogo;NSP;NSP-CL;r;Reticulon-4;Rtn4;RTN4;RTN4-A;RTN4-B1;RTN4-B2;RTN4-C;RTN-X
• NI220
• NogoA
• Nogo-A
• RTN4
• RTN4A

Background

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.

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.

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-A Antibody (AF3515)(4)

Publications using AF3515

• Y Sekine, JA Lindborg, SM Strittmatt A proteolytic C-terminal fragment of Nogo-A (reticulon-4A) is released in exosomes and potently inhibits axon regeneration Nat. Med., 2019-11-20;295(8):2175-2183. 2019-11-20 [PMID: 31748413] (Western Blot, Human)
• Y Chen, W Zhen, T Guo, Y Zhao, A Liu, JP Rubio, D Krull, JC Richardson, H Lu, R Wang Histamine Receptor 3 negatively regulates oligodendrocyte differentiation and remyelination PLoS ONE, 2017-12-18;12(12):e0189380. 2017-12-18 [PMID: 29253893] (IHC-Fr, Mouse)
• Jae Young Lee, Min Joung Kim, Lijun Li, Alexander A. Velumian, Pei Mun Aui, Michael G. Fehlings et al. Nogo receptor 1 regulates Caspr distribution at axo-glial units in the central nervous system Scientific Reports 8/21/2017 [PMID: 28827698]
• Sumiyoshi K, Obayashi S, Tabunoki H Protein microarray analysis identifies cyclic nucleotide phosphodiesterase as an interactor of Nogo-A. Neuropathology, 2009-06-07;30(1):7-14. 2009-06-07 [PMID: 19508346] (IHC-P, Western Blot, Mouse, Rat)

Bioinformatics

• Gene Symbol: RTN4
• Uniprot:
  • Q9NQC3()