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RecombinantRabMAb

Recombinant Anti-Nav1.6/SCN8A antibody [EPR25137-45] (ab302786)

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Western blot - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Western blot - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunoprecipitation - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Immunoprecipitation - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
  • Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25137-45] to Nav1.6/SCN8A
  • Suitable for: WB, IHC-Fr, IP, IHC-P
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Nav1.6/SCN8A antibody [EPR25137-45]
    See all Nav1.6/SCN8A primary antibodies

  • Description

    Rabbit monoclonal [EPR25137-45] to Nav1.6/SCN8A

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Fr, IP, IHC-Pmore details
    Unsuitable for: Flow Cyt (Intra) or ICC/IF

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Mouse brain tissue lysate. Rat spinal cord tissue lysate. Human cerebellum and spinal cord tissue lysate. HEK-293T cells transfected with a his tagged Human SCN8A expression vector, whole cell lysate. IHC-P: Mouse and rat cerebrum tissue. IHC-Fr: Mouse and rat frozen cerebrum tissue (fresh). IP: Mouse and rat brain tissue lysate.

  • General notes

    ab302786 does not react in: IHC-P with human samples; ICC/IF with human, mouse, and rat species; intracellular flow cytometry with mouse and rat samples. 

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab302786 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
1/1000. Detects a band of approximately 260 kDa (predicted molecular weight: 225 kDa).
IHC-Fr
1/500.
IP
1/30.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/1000. Detects a band of approximately 260 kDa (predicted molecular weight: 225 kDa).
IHC-Fr
1/500.
IP
1/30.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for Flow Cyt (Intra) or ICC/IF.

Target

  • Function

    Mediates the voltage-dependent sodium ion permeability of excitable membranes. Assuming opened or closed conformations in response to the voltage difference across the membrane, the protein forms a sodium-selective channel through which Na(+) ions may pass in accordance with their electrochemical gradient. In macrophages and melanoma cells, isoform 5 may participate in the control of podosome and invadopodia formation.

  • Tissue specificity

    Isoform 5 is expressed in non-neuronal tissues, such as monocytes/macrophages.

  • Sequence similarities

    Belongs to the sodium channel (TC 1.A.1.10) family. Nav1.6/SCN8A subfamily.
    Contains 1 IQ domain.

  • Domain

    The sequence contains 4 internal repeats, each with 5 hydrophobic segments (S1,S2,S3,S5,S6) and one positively charged segment (S4). Segments S4 are probably the voltage-sensors and are characterized by a series of positively charged amino acids at every third position.

  • Post-translational
    modifications

    May be ubiquitinated by NEDD4L; which would promote its endocytosis.

  • Cellular localization

    Membrane and Cytoplasmic vesicle. Some vesicles are localized adjacent to melanoma invadopodia and macrophage podosomes. Does not localize to the plasma membrane.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • CerIII antibody
    • CIAT antibody
    • EIEE13 antibody
    • hNa6/Scn8a voltage gated sodium channel antibody
    • MED antibody
    • Motor endplate disease antibody
    • NaCh 6 antibody
    • NaCh6 antibody
    • Nav 1.6 antibody
    • Nbna1 antibody
    • peripheral nerve protein type 4 antibody
    • PN 4 antibody
    • PN4 antibody
    • SCN8A antibody
    • SCN8A_HUMAN antibody
    • Sodium channel protein type 8 alpha subunit antibody
    • Sodium channel protein type 8 subunit alpha antibody
    • Sodium channel protein type VIII alpha subunit antibody
    • Sodium channel protein type VIII subunit alpha antibody
    • Sodium channel voltage gated type VIII alpha antibody
    • Sodium channel voltage gated type VIII alpha polypeptide antibody
    • Sodium channel voltage gated type VIII alpha subunit antibody
    • Voltage gated sodium channel subunit alpha Nav1.6 antibody
    • Voltage-gated sodium channel subunit alpha Nav1.6 antibody
    see all

Images

  • Western blot - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Western blot - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    All lanes : Anti-Nav1.6/SCN8A antibody [EPR25137-45] (ab302786) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Mouse liver tissue lysate
    Lane 3 : Rat spinal cord tissue lysate
    Lane 4 : Rat liver tissue lysate
    Lane 5 : Human spinal cord tissue lysate
    Lane 6 : Human cerebellum tissue lysate
    Lane 7 : Human liver tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Performed under non-reducing conditions.

    Predicted band size: 225 kDa
    Observed band size: 260 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    The expression molecular weight observed is consistent with what has been described in the literature (PMID: 26410685).

    Negative control: Liver (PMID: 11532991).

    Bands around 150 kDa and 250 kDa may due to degradation.

    Samples are non-boiled as boiling may cause protein aggregates.
    Lane 5-7 of the blot was developed using a high sensitivity ECL substrate. 

    Exposure time: Lane 1-4: 114 seconds, Lane 5-7: 48 seconds.

  • Western blot - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Western blot - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    All lanes : Anti-Nav1.6/SCN8A antibody [EPR25137-45] (ab302786) at 1/1000 dilution

    Lane 1 : HEK-293T cells transfected with a Human SCN8A expression vector containing a his tag, whole cell lysate
    Lane 2 : HEK-293T cells transfected with a Human SCN2A expression vector containing a his tag, whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Performed under non-reducing conditions.

    Predicted band size: 225 kDa
    Observed band size: 260 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

    Samples are non-boiled as boiling may cause protein aggregates.
    This antibody does not cross-react with human SCN2A.

    Exposure time: 3.25 seconds. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling Nav1.6/SCN8A with ab302786 at 1/2000 dilution (0.246 µg/ml), followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on axon in mouse cerebrum. The section was incubated with ab302786 at 4°C overnight. Counterstained with Hematoxylin.

    Secondary antibody only control: A ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of paraffin-embedded rat cerebrum tissue labeling Nav1.6/SCN8A with ab302786 at 1/2000 dilution (0.246 µg/ml), followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). Positive staining on axon in rat cerebrum. The section was incubated with ab302786 at 4°C overnight. Counterstained with Hematoxylin.

    Secondary antibody only control: A ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Nav1.6/SCN8A with ab302786 at 1/2000 dilution (0.246 µg/ml), followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). The section was incubated with ab302786 at 4°C overnight. Counterstained with Hematoxylin.

    Negative control: No staining on mouse liver is observed. 

    Secondary antibody only control: A ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Nav1.6/SCN8A with ab302786 at 1/2000 dilution (0.246 µg/ml), followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer). The section was incubated with ab302786 at 4°C overnight. Counterstained with Hematoxylin.

    Negative control: No staining on rat liver is observed. 

    Secondary antibody only control: A ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).

    Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum (fresh) tissue labeling Nav1.6/SCN8A with ab302786 at 1/500 dilution (0.982 µg/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/ml) (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).

    Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 µg/ml). 

  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labeling Nav1.6/SCN8A with ab302786 at 1/500 dilution (0.982 µg/ml), followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

    Negative control: no staining on mouse liver is observed (PMID: 11532991). 

    Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution. 

  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cerebrum (fresh) tissue labeling Nav1.6/SCN8A with ab302786 at 1/500 (0.982 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

    Positive staining on rat cerebrum is observed.

    Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution. 

  • Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunohistochemistry (Frozen sections) - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labeling Nav1.6/SCN8A with ab302786 at 1/500 (0.982 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

    Negative control: no staining on rat liver is observed (PMID: 11532991).

    Secondary antibody control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 µg/ml) dilution. 

  • Immunoprecipitation - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunoprecipitation - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Nav1.6/SCN8A was immunoprecipitated from 0.35 mg mouse brain tissue lysate with ab302786 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302786 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: mouse brain tissue lysate 10 µg (Input)

    Lane 2: ab302786 IP in mouse brain tissue lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302786 in mouse brain tissue lysate

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 15 seconds.

    Observed MW (kDa): 260

    Input is non-boiled as boiling may cause protein aggregates.

  • Immunoprecipitation - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Immunoprecipitation - Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

    Nav1.6/SCN8A was immunoprecipitated from 0.35 mg rat brain tissue lysate with ab302786 at 1/30 dilution (2 µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302786 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: rat brain tissue lysate 10 µg (Input)

    Lane 2: ab302786 IP in rat brain tissue lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab302786 in rat brain tissue lysate

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 15 seconds.

    Observed MW (kDa): 260 

    Input is non-boiled as boiling may cause protein aggregates.

  • Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)
    Anti-Nav1.6/SCN8A antibody [EPR25137-45] (AB302786)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab302786? Please let us know so that we can cite the reference in this datasheet.

ab302786 has not yet been referenced specifically in any publications.

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