Product nameAnti-Neurofascin antibody
See all Neurofascin primary antibodies
DescriptionRabbit polyclonal to Neurofascin
Tested applicationsSuitable for: WB, ICC/IF, IHC-P, IHC-FoFrmore details
Species reactivityReacts with: Mouse, Rat, Human, Common marmoset
- This antibody gave a positive signal in the following lysates; Brain (Mouse) Tissue Lysate - normal tissue Brain (Rat) Tissue Lysate - normal tissue
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab31457 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 155,186 kDa (predicted molecular weight: 138 kDa).|
|ICC/IF||Use a concentration of 1 µg/ml.|
|IHC-P||1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
FunctionCell adhesion, ankyrin-binding protein which may be involved in neurite extension, axonal guidance, synaptogenesis, myelination and neuron-glial cell interactions.
Sequence similaritiesBelongs to the immunoglobulin superfamily. L1/neurofascin/NgCAM family.
Contains 5 fibronectin type-III domains.
Contains 6 Ig-like C2-type (immunoglobulin-like) domains.
Cellular localizationCell membrane.
- Information by UniProt
- KIAA0756 antibody
- Neurofascin antibody
- Neurofascin homolog antibody
Anti-Neurofascin antibody (ab31457) at 1 µg/ml + Brain(Mouse)Whole Cell Lysate at 10 µg
IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 138 kDa
Observed band size: 155,186 kDa why is the actual band size different from the predicted?
Additional bands at: 120 kDa, 55 kDa. We are unsure as to the identity of these extra bands.In Rat, Neurofascin has three observed isoforms , isoform 1 (138 kDa) which is known as Nfasc186, isoform 2 (133 kDa) and isoform 3 (132 kDa) which are both known as Nfasc155. These proteins are highly glycosylated and their observed molecular weight in Western blot is 186 kDa and 155 kDa respectively.
Mouse Neurofascin has a predicted molecular weight of 138 kDa (Swiss-Prot data) however the observed bands at 186 kDa and 155 kDa are thought to correspond to the glycosylated isoforms 1, 2 and 3 as observed in Rat (Sherman DL et al, 2005,PubMed:16337912)
ICC/IF image of ab31457 stained human SHSY5Y cells. The cells were 4% PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31475, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
Ab31457 used on Rat cerebellum, Purkinje cells are variously positive: there is a clear punctate positivity of secondary dendritic processes, a strong membraneous positivity of the somata and also a single strongly positive fibre that seemingly enters the Granule Cell Layer indicated by red arrowheads axons of P.cells. The Glomeruli of the GCL are moderately positive. There is also a linear pattern of positivity within the GCL that may well be fibres.
Immunohistochemistical detection of Neurofascin using antibody ab31457 on PFA perfusion-fixed rat brain sections. Primary antibody ab31457 was used at 1/1000 incubated for 18 hours @ 20°C in PBS + 0.3 % Triton X100. Secondary antibody: goat anti-rabbit Alexa Fluor 488 conjugated (1/1000). Immunostaining obtained in the rat hippocampus using this antibody diluted 1:1000. Sections come from a perfused-fixed animal. The immunostaining was performed using the `free floating` technique, using direct fluorescence. Some cytoplasmic staining was observed in granular neurons and neuronal processes (arrows on the figure on the right). The lefthand figure shows the hippocampal granular cell staining (objective X20), the righthand figure shows this at higher magnification. This staining is consistent with published observation of cytoplasmic localisation of Neurofascin in the cytoplasm of hippocampal cells and their axon initial segment (Burkarth
This product has been referenced in:
- Monfrini E et al. Neurofascin (NFASC) gene mutation causes autosomal recessive ataxia with demyelinating neuropathy. Parkinsonism Relat Disord N/A:N/A (2019). Read more (PubMed: 30850329) »
- Edamakanti CR et al. Mutant ataxin1 disrupts cerebellar development in spinocerebellar ataxia type 1. J Clin Invest 128:2252-2265 (2018). Read more (PubMed: 29533923) »