Anti-Neurofascin antibody (ab31457)
Key features and details
- Rabbit polyclonal to Neurofascin
- Suitable for: WB, ICC/IF, IHC-FoFr
- Reacts with: Mouse, Rat, Human, Common marmoset
- Isotype: IgG
Overview
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Product name
Anti-Neurofascin antibody
See all Neurofascin primary antibodies -
Description
Rabbit polyclonal to Neurofascin -
Host species
Rabbit -
Specificity
The ab31457 antibody would be able to bind to the full length canonical sequence Isoform 1 and also to Isoforms 5, Isoform 6, Isoform 9 and Isoform 13.
The antibody would not be able to detect Isoforms 2, 3, 4, 7, 8, 10, 11 or Isoform 12. All of these Isoforms are missing amino acids 1035-1203. -
Tested applications
Suitable for: WB, ICC/IF, IHC-FoFrmore details -
Species reactivity
Reacts with: Mouse, Rat, Human, Common marmoset -
Immunogen
Properties
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Form
Liquid -
Storage instructions
Shipped 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 buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
Applications
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.
Application Abreviews Notes 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-FoFr 1/1000. Target
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Function
Cell adhesion, ankyrin-binding protein which may be involved in neurite extension, axonal guidance, synaptogenesis, myelination and neuron-glial cell interactions. -
Sequence similarities
Belongs to the immunoglobulin superfamily. L1/neurofascin/NgCAM family.
Contains 5 fibronectin type-III domains.
Contains 6 Ig-like C2-type (immunoglobulin-like) domains. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 23114 Human
- Entrez Gene: 269116 Mouse
- Entrez Gene: 116690 Rat
- Omim: 609145 Human
- SwissProt: O94856 Human
- SwissProt: Q810U3 Mouse
- SwissProt: P97685 Rat
- Unigene: 13349 Human
see all -
Alternative names
- KIAA0756 antibody
- Neurofascin antibody
- Neurofascin homolog antibody
see all
Images
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Western blot - Anti-Neurofascin antibody (ab31457)Anti-Neurofascin antibody (ab31457) at 1 µg/ml + Brain(Mouse)Whole Cell Lysate at 10 µg
Secondary
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) -
Immunocytochemistry/ Immunofluorescence - Anti-Neurofascin antibody (ab31457)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). -
Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Neurofascin antibody (ab31457)This image is courtesy of Sophie Pezet, CNRS, Paris, FranceImmunohistochemistical 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
Protocols
Datasheets and documents
References (16)
ab31457 has been referenced in 16 publications.
- Monfrini E et al. Neurofascin (NFASC) gene mutation causes autosomal recessive ataxia with demyelinating neuropathy. Parkinsonism Relat Disord N/A:N/A (2019). PubMed: 30850329
- Edamakanti CR et al. Mutant ataxin1 disrupts cerebellar development in spinocerebellar ataxia type 1. J Clin Invest 128:2252-2265 (2018). PubMed: 29533923
- Kanellopoulos AH et al. Mapping protein interactions of sodium channel NaV1.7 using epitope-tagged gene-targeted mice. EMBO J 37:427-445 (2018). PubMed: 29335280
- Suzuki S et al. Spatio-temporal and dynamic regulation of neurofascin alternative splicing in mouse cerebellar neurons. Sci Rep 7:11405 (2017). PubMed: 28900163
- D'Este E et al. Ultrastructural anatomy of nodes of Ranvier in the peripheral nervous system as revealed by STED microscopy. Proc Natl Acad Sci U S A 114:E191-E199 (2017). PubMed: 28003466
Images
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Western blot - Anti-Neurofascin antibody (ab31457)Anti-Neurofascin antibody (ab31457) at 1 µg/ml + Brain(Mouse)Whole Cell Lysate at 10 µg
Secondary
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) -
Immunocytochemistry/ Immunofluorescence - Anti-Neurofascin antibody (ab31457)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).
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Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-Neurofascin antibody (ab31457) This image is courtesy of Sophie Pezet, CNRS, Paris, France
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
