Neurofascin Antibody

Western blot shows lysates of rat brain tissue, mouse brain tissue, and human brain (cerebellum). PVDF Membrane was probed with 0.1 µg/mL of Chicken Anti-Human/Mouse/Rat Neurofascin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3235) followed by HRP-conjugated Anti-Chicken IgY Secondary Antibody. Specific bands were detected for Neurofascin at approximately 140 and186 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Neurofascin was detected in perfusion fixed frozen sections of rat brain (dorsal root ganglion) using Chicken Anti-Human/Mouse/Rat Neurofascin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3235) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Chicken IgY Secondary Antibody (yellow; Catalog # NL016) and counterstained with DAPI (blue). Specific staining was localized to neuronal cell bodies and Schwann cells (perinodal regions). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

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

Neurofascin Antibody Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant rat Neurofascin
  Ile25-Ala1031
  Accession # NP_446361
• Specificity: Detects human, mouse, rat Neurofascin in Western blots and rat Neurofascin in direct ELISAs.
• Source: N/A
• Isotype: IgY
• Clonality: Polyclonal
• Host: Chicken
• Gene: NFASC
• Purity: Immunogen affinity purified
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 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 °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
• Purity: Immunogen affinity purified
• 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 Neurofascin Antibody

• DKFZp686P2250
• KIAA0756FLJ46866
• neurofascin homolog (chicken)
• neurofascin homolog
• Neurofascin
• NF
• NFASC
• NRCAML

Background

Neurofascin 155 (NF155) is a type I transmembrane glycoprotein that belongs to the L1CAM family of cell adhesion proteins (1, 2). The rat NF155 cDNA encodes a 1240 amino acid (aa) precursor that contains a 24 aa signal sequence, a 1086 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 109 aa cytoplasmic domain. The ECD consists of six Ig-like domains and four fibronectin type III repeats, the second of which has an RGD motif. A splice variant of Neurofascin, NF186, lacks the RGD-containing fibronectin type III domain but instead has a mucin-like domain and an additional non-RGD fibronectin type III domain (3). Within shared regions of the ECD, rat NF155 shares 45% and 39% aa sequence identity with rat Nr-CAM and L1CAM, respectively, and 98% aa sequence identity with human and mouse NF155. NF155 is transiently expressed by oligodendrocytes at the onset of axon myelination, whereas NF186 is neuronally expressed in nodes of Ranvier (4‑6). Clustering of NF155 in paranodal oligodengroglia lipid raft domains is stabilized by dimerization of its cytoplasmic domains and association with intracellular ankyrin (6‑9). NF155 interacts with axonal contactin and plays a role in node of Ranvier formation and the establishment of saltatory conduction (5, 9‑12). The ECD of NF155 is cleaved from oligodengroglia membranes by metalloproteases, a process which is required for NF155 transport from the glial cell body to the axoglial junction (13). In addition to distinct expression patterns, Neurofascin isoforms have different functional properties. NF155 promotes neuronal adhesion and neurite outgrowth, whereas NF186 inhibits neuronal adhesion (4, 7, 13).

1. Sherman, D.L. and P.J. Brophy (2005) Nat. Rev. Neurosci. 6:683.
2. Coman, I. et al. (2005) J. Neurol. Sci. 233:67.
3. Volkmer, H. et al. (1992) J. Cell Biol. 118:149.
4. Koticha, D. et al. (2005) Mol. Cell. Neurosci. 30:137.
5. Collinson, J.M. et al. (1998) Glia 23:11.
6. Schafer, D.P. et al. (2004) J. Neurosci. 24:3176.
7. Maier, O. et al. (2005) Mol. Cell. Neurosci. 28:390.
8. Zhang, X. et al. (1998) J. Biol. Chem. 273:30785.
9. Sherman, D.L. et al. (2005) Neuron 48:737.
10. Gollan, L. et al. (2003) J. Cell Biol. 163:1213.
11. Charles, P. et al. (2002) Curr. Biol. 12:217.
12. Volkmer, H. et al. (1996) J. Cell Biol. 135:1059.
13. Maier, O. et al. (2006) Exp. Cell Res. 312:500.

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 Neurofascin Antibody (AF3235)(10)

Publications using AF3235

• JA Miller, DE Drouet, LM Yermakov, MS Elbasiouny, FZ Bensabeur, M Bottomley, K Susuki Distinct Changes in Calpain and Calpastatin during PNS Myelination and Demyelination in Rodent Models International Journal of Molecular Sciences, 2022;23(23):. 2022 [PMID: 36499770] (IHC, Rat)
• G Shacklefor, LN Marziali, Y Sasaki, A Claessens, C Ferri, NI Weinstock, AM Rossor, NJ Silvestri, ER Wilson, E Hurley, GJ Kidd, S Manohar, D Ding, RJ Salvi, ML Feltri, M D'Antonio, L Wrabetz A new mouse model of Charcot-Marie-Tooth 2J neuropathy replicates human axonopathy and suggest alteration in axo-glia communication PloS Genetics, 2022;18(11):e1010477. 2022 [PMID: 36350884] (IHC, Mouse)
• G Della-Flor, ER Wilson, LN Marziali, E Hurley, N Silvestri, B He, BW O'Malley, B Beirowski, Y Poitelon, L Wrabetz, ML Feltri Prohibitin 1 is essential to preserve mitochondria and myelin integrity in Schwann cells Nature Communications, 2021;12(1):3285. 2021 [PMID: 34078899] (IHC, Mouse)
• KJ Chang, I Agrawal, A Vainshtein, WY Ho, W Xin, G Tucker-Kel, K Susuki, E Peles, SC Ling, JR Chan TDP-43 maximizes nerve conduction velocity by repressing a cryptic exon for paranodal junction assembly in Schwann cells Elife, 2021;10(0):. 2021 [PMID: 33689679] (IHC, Mouse)
• AJ Davies, J Fehmi, M Senel, H Tumani, J Dorst, S Rinaldi Immunoadsorption and Plasma Exchange in Seropositive and Seronegative Immune-Mediated Neuropathies J Clin Med, 2020;9(7):. 2020 [PMID: 32605107] (Bioassay, Human)
• Y Zhang, S Yuen, E Peles, JL Salzer Accumulation of Neurofascin at nodes of Ranvier is regulated by a Paranodal Switch J. Neurosci., 2020;0(0):. 2020 [PMID: 32554548] (Western Blot, Transgenic Mouse)
• K Susuki, DR Zollinger, KJ Chang, C Zhang, CY Huang, CR Tsai, MR Galiano, Y Liu, SD Benusa, LM Yermakov, RB Griggs, JL Dupree, MN Rasband Glial ?II spectrin contributes to paranode formation and maintenance J. Neurosci., 2018;0(0):. 2018 [PMID: 29853631] (IHC, Mouse)
• RB Griggs, LM Yermakov, DE Drouet, DVM Nguyen, K Susuki Methylglyoxal Disrupts Paranodal Axoglial Junctions via Calpain Activation ASN Neuro, 2018;10(0):1759091418766. 2018 [PMID: 29673258] (IHC, Mouse)
• L Querol, AM Siles, R Alba-Rovir, A Jáuregui, J Devaux, C Faivre-Sar, J Araque, R Rojas-Garc, J Diaz-Maner, E Cortés-Vic, G Nogales-Ga, M Navas-Madr, E Gallardo, I Illa Antibodies against peripheral nerve antigens in chronic inflammatory demyelinating polyradiculoneuropathy Sci Rep, 2017;7(1):14411. 2017 [PMID: 29089585] (ICC, Human)
• V Amor, C Zhang, A Vainshtein, A Zhang, DR Zollinger, Y Eshed-Eise, PJ Brophy, MN Rasband, E Peles The paranodal cytoskeleton clusters Na(+) channels at nodes of Ranvier Elife, 2017;6(0):. 2017 [PMID: 28134616] (IHC, Mouse)

Review for Neurofascin Antibody (AF3235) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Verified Customer IF Human 07/16/2015

Summary

• Application: Immunofluorescence
• Sample Tested: HEK 293 cell line transfected with Neurofascin expression plasmid
• Species: Human
• Lot: CFCT0214081

FAQs for Neurofascin Antibody (AF3235). (Showing 1 - 1 of 1 FAQs).

1. Does the neurofascin antibody #AF3235 detect both 155 and 186 isoforms?
   • The immunogen for this antibody is "Mouse myeloma cell line NS0-derived recombinant rat Neurofascin Ile25 - Ala1031". It means that any protein target has the the partial or full-length region that is included in Ile25 - Ala1031 will be recognized because it is a polyclonal antibody. Then, I checked the rat sequence posting on UniProt (http://www.uniprot.org/uniprot/P97685#sequences), it says that rat has three Neurofascin isoforms: 1 to 3. Isoform has 1,240 a.a. with a mass of 138,004 (Da), whereas both isoforms 2 and 3 are known as "NF155, 155 kDa isoforms" According above information, the isoforms 2 and 3 should also be recognized by the antibody, AF3235, because the missing and sequence-substituted regions are not excluded out of "Ile25 - Ala1031"; and there are still a lot of epitopes left on the protein species for binding. Unfortunately, I did not locate any information on isoform 186 (assuming this isoform has ~186 kDa) in rat, human Uniprot, or mouse Uniprot databases. If you have the a.a. sequence for the isoform, you could analyze the structure and determine whether AF3235 can recognize it.

Bioinformatics

• Gene Symbol: NFASC
• Uniprot:
  • NP_446361(Human)