Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat, Porcine, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot

Cited:

Immunohistochemistry, Western Blot, Immunocytochemistry, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Chicken IgY
View all Neurofascin Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Chicken

Isotype

IgY

Scientific Data Images for Human/Mouse/Rat Neurofascin Antibody

Detection of Human, Mouse, and Rat Neurofascin antibody by Western Blot.

Detection of Human, Mouse, and Rat Neurofascin by Western Blot.

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 antibody in Rat Brain by Immunohistochemistry (IHC-Fr).

Neurofascin in Rat Brain.

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.
Detection of Mouse Neurofascin by Immunohistochemistry

Detection of Mouse Neurofascin by Immunohistochemistry

P0T124M mutation alters SLI morphology, length,&distribution. (D) Quantification shows an increased %age of SLIs in nerves from mice harboring the P0T124M mutation (MpzT124M) at 2, 6, 12,&18 months of age. One-way ANOVA [2 month old: F (2, 10) = 29.86, p<0.0001; 6 month old: F (2, 9) = 41.60, p<0.0001, 12 month old: F (2, 6) = 34.35, p = 0.0005; 18 month old: F (2, 8) = 21.71, p = 0.0006]. Representative confocal pictures of sciatic teased fibers stained with FITC-phalloidin (ACTIN, green) (E, I,&M) or anti-MAG antibodies (Q) (green)&anti-pan-NFASC antibodies (red) at 2 (E), 6 (I),&12 (M&Q) months of age. SLI morphology is disrupted in MpzT124M mice. Scale bar: 10 μm. Measurements of SLI length at 2 (F), 6 (J),&12 (N&R) months of age. Nested one-way ANOVA [2 month old: F (2,9) = 32.16, p <0.0001; 6 month old: F (2,6) = 11.18, p = 0.0095 12 month old: F (2,9) = 6.447, p = 0.0183]. Measurements of the distance between adjacent SLI at 2 (G), 6 (K),&12 (O&S) months of age. Nested one-way ANOVA [2 month old: F (2,9) = 24.90, p = 0.0002; 6 month old: F (2,6) = 9.382, p = 0.0142; 12 month old: F (2,9) = 13.04, p = 0.0022]. Quantifications of SLI number per 100 μm at 2 (H), 6 (L),&12 (P&T) months of age. Nested one-way ANOVA [2 month old: F (2,9) = 20.29, p = 0.0005; 6 month old: F (2,6) = 22.81, p = 0.016; 12 month old: F (2,9) = 28.25, p = 0.0001]. At least 207 SLI per genotype quantified at each time point. n (animals) ≥ 3 per genotype. *p < 0.05, **p < 0.01, ***p < 0.001 by multiple-comparisons Tukey’s post hoc tests after Nested one-way ANOVA (D, F, G, H, J, K, L, R, S,&T) or by two-tailed Student’s t test (N, O,&P). Graphs indicate means ± SEMs. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36350884), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Neurofascin by Immunohistochemistry

Detection of Mouse Neurofascin by Immunohistochemistry

P0T124M mutation alters nodes and paranodes.Representative confocal pictures of sciatic nerve teased fibers stained with antibodies against the paranodal marker CASPR (red) and the paranodal and nodal marker pan-Neurofascin (NFASC; green) at 2 (A) and 12 (E) months of age. Scale bars: 5 μm. CASPR length quantifications at 2 (B) and 12 (F) months of age. Nested one-way ANOVA [2 month old: F (2,9) = 7.009, p = 0.0146; 12 month old: F (2,7) = 35.47, p = 0.0002]. Relative frequency distributions of paranodal (CASPR) length at 2 (C) and 12 (G) months of age. Nodal length quantifications at 2 (D) and 12 (H) months of age. Nested one-way ANOVA [2 month old: F (2,9) = 1.122, p = 0.3671, 12 month old: F (2,7) = 8.012, p = 0.0155]. At least 200 paranodes and 100 nodes per genotype were quantified at each time point. n (animals) ≥ 3 per genotype. (I) Electron micrographs of ultrathin longitudinal WT and MpzT124M/T124M sciatic nerve section at 12 months of age. In (I) electron micrographs represent nodes of WT and MpzT124M/T124M sciatic nerves. (J) The quantification of nodal length shows significant widening of the node in MpzT124M/T124M. Magnifications of (I) show disorganized paranonal loops in MpzT124M/T124M but well organized in WT. n (animals) ≥ 3 per genotype, 7 to 11 nodes were counted per animals, for a total of 27 and 28 counted by genotype. Scale bare: top panel 1 μm, middle panel 500 nm, bottom panel 200 nm. ***p < 0.001 by multiple-comparisons Tukey’s post hoc tests after Nested one-way ANOVA (B, F, D, and H) or Nested two-tailed Student’s t test (J). Graphs indicate means ± SEMs. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36350884), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat Neurofascin Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample: Perfusion fixed frozen sections of rat brain (dorsal root ganglion)

Western Blot

0.1 µg/mL
Sample: Rat brain tissue, mouse brain tissue, and human brain (cerebellum) tissue

Reviewed Applications

Read 1 review rated 5 using AF3235 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Neurofascin

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).

References

  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.

Alternate Names

NFASC, NRCAML

Entrez Gene IDs

23114 (Human); 269116 (Mouse); 116690 (Rat)

Gene Symbol

NFASC

UniProt

NP_446361 (Human)

Additional Neurofascin Products

Product Documents for Human/Mouse/Rat Neurofascin Antibody

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Product Specific Notices for Human/Mouse/Rat Neurofascin Antibody

For research use only

Citations for Human/Mouse/Rat Neurofascin Antibody

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  • Name: YONGJIE LAI
    Application: Immunofluorescence
    Sample Tested: HEK 293 cell line transfected with Neurofascin expression plasmid
    Species: Human
    Verified Customer | Posted 07/16/2015

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FAQs for Human/Mouse/Rat Neurofascin Antibody

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  • Q: Does the neurofascin antibody #AF3235 detect both 155 and 186 isoforms?

    A:

    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.

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