製品の概要

  • 製品名
    Anti-Nav1.7 antibody [N68/6]
    Nav1.7 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [N68/6] to Nav1.7
  • 由来種
    Mouse
  • 特異性
    No cross reactivity against other Nav channels.
  • アプリケーション
    適用あり: WB, ICC/IF, IP, IHC-P, IHC-Fr, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Fusion protein, corresponding to cytoplasmic C terminal amino acids 1751-1946 of Human Nav1.7.

  • ポジティブ･コントロール
    • HEK-293 cell lysate transiently expressing Nav1.7
  • 特記事項

    The clone number has been updated from S68-6 to N68/6, both clone numbers name the same antibody clone.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab85015 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 - 10 µg/ml. Predicted molecular weight: 226 kDa.
ICC/IF Use a concentration of 1 - 10 µg/ml.
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 0.1 - 1 µg/ml.
IHC-Fr Use a concentration of 0.1 - 1 µg/ml.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ターゲット情報

  • 機能
    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 (PubMed:7720699, PubMed:17167479, PubMed:25240195, PubMed:26680203, PubMed:15385606, PubMed:16988069, PubMed:17145499, PubMed:19369487, PubMed:24311784). It is a tetrodotoxin-sensitive Na(+) channel isoform (PubMed:7720699). Plays a role in pain mechanisms, especially in the development of inflammatory pain (PubMed:17167479, PubMed:17145499, PubMed:19369487, PubMed:24311784).
  • 組織特異性
    Expressed strongly in dorsal root ganglion, with only minor levels elsewhere in the body, smooth muscle cells, MTC cell line and C-cell carcinoma. Isoform 1 is expressed preferentially in the central and peripheral nervous system. Isoform 2 is expressed preferentially in the dorsal root ganglion.
  • 関連疾患
    Primary erythermalgia
    Indifference to pain, congenital, autosomal recessive
    Paroxysmal extreme pain disorder
    Generalized epilepsy with febrile seizures plus 7
    Febrile seizures, familial, 3B
  • 配列類似性
    Belongs to the sodium channel (TC 1.A.1.10) family. Nav1.7/SCN9A subfamily.
    Contains 1 IQ domain.
  • ドメイン
    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.
  • 翻訳後修飾
    Phosphorylation at Ser-1490 by PKC in a highly conserved cytoplasmic loop increases peak sodium currents.
    Ubiquitinated by NEDD4L; which may promote its endocytosis. Does not seem to be ubiquitinated by NEDD4.
  • 細胞内局在
    Cell membrane. Cell projection. In neurite terminals.
  • Information by UniProt
  • 参照データベース
    see all
  • 別名
    • ETHA antibody
    • GEFSP7 antibody
    • hNE Na antibody
    • hNE-Na antibody
    • hNENa antibody
    • NE NA antibody
    • NENA antibody
    • Neuroendocrine sodium channel antibody
    • Peripheral sodium channel 1 antibody
    • PN1 antibody
    • Scn9a antibody
    • SCN9A_HUMAN antibody
    • Sodium channel protein type 9 subunit alpha antibody
    • Sodium channel protein type IX subunit alpha antibody
    • Sodium channel voltage gated type IX alpha antibody
    • Sodium channel voltage gated type IX alpha polypeptide antibody
    • Sodium channel voltage gated type IX alpha subunit antibody
    • Voltage gated sodium channel alpha subunit Nav1.7 antibody
    • Voltage gated sodium channel subunit alpha Nav1 antibody
    • Voltage-gated sodium channel subunit alpha Nav1.7 antibody
    see all

画像

  • Immunohistochemistry - Anti-Nav1.7 antibody [N68/6] (ab85015)
    Immunohistochemistry - Anti-Nav1.7 antibody [N68/6] (ab85015)

     Immunohistochemical analysis of Mouse brain tissue labeling Nav1.7 with ab85015 at 1μg/ml.

  • Western blot - Anti-Nav1.7 antibody [N68/6] (ab85015)
    Western blot - Anti-Nav1.7 antibody [N68/6] (ab85015)
    All lanes : Anti-Nav1.7 antibody [N68/6] (ab85015) at 1 µg/ml

    Lane 1 : Molecular weight marker
    Lane 2 : Cell lysates prepared from CHO cell line constitutively expressing Nav1.7 at 10 µg

    Predicted band size: 226 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.7 antibody [N68/6] (ab85015)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nav1.7 antibody [N68/6] (ab85015)

    ab85015 at 1/100 dilution staining Nav1.7 in mouse back skin tissue section by IHC-P. Bouin's fixed and paraffin-embedded tissue sections were used. Tissue underwent heat mediated antigen retrieval in microwave with two, 5 minutes incubation intervals in citrate buffer. A Fluorophore conjugated goat anti mouse at 1/50 dilution was used as secondary.

  • Flow Cytometry - Anti-Nav1.7 antibody [N68/6] (ab85015)
    Flow Cytometry - Anti-Nav1.7 antibody [N68/6] (ab85015)
    Overlay histogram showing SH-SY5Y cells stained with ab85015 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab85015, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive result in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween for 20 min used under the same conditions.

参考文献

This product has been referenced in:
  • Bi RY  et al. Estradiol upregulates voltage-gated sodium channel 1.7 in trigeminal ganglion contributing to hyperalgesia of inflamed TMJ. PLoS One 12:e0178589 (2017). IHC-Fr ; Rat . Read more (PubMed: 28582470) »

See 1 Publication for this product

ab85015 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

レビューと Q&A

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12 reviews or Q&A

10
Votes: Highest First

Immunocytochemistry/ Immunofluorescence abreview for Anti-Nav1.7 antibody [S68-6]

 Good
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (NSC derived neurons)
Permeabilization
Yes - Triton X-100
Specification
NSC derived neurons
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Paraformaldehyde
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abreview image
Username

Mr. Thomas Klein

Verified customer

投稿 Nov 13 2015

How was non-crossreactivity with other Nav channels of both of these antibodies determined?

The specificity of these antibodies to their target Nav channel was determined by performing western blot analysis on lysates from various HEK cell lines expressing one Nav channel. These antibodies were used to detect the presence of the channel in th...

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Please find below the details about our Western blot protocol that we used to detect Nav1.7 with you antibody [S68-6] (ab85015).

We are looking forward for your comments.



Sample (species) and full or fragment: extracts...

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Thank you for your response and for taking the time for providing some useful information.

After reading through the detailed protocol you kindly forwarded to Abcam, I would like to make the following comments/suggestions:

Transfect...

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Dear,
Thank you. I certainly will take your suggestion into account. I also like to point out that our WB is dry (7 min transfer, although I've treid 14 min), so no membrane manipulation is possible
Thanks

Thank you for getting back to me.
although it may be beneficial to optimise the transfer as far as possible (and I would still suggest staining the membrane to observe the protein transfer if possible), you have seen some signal within the expecte...

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Dear,
First of all, thank you for your effort. As an answer to your questions, please see below:

I'm please do hear that the antibody has been working well in flow cytometry. It is unfortunate that the Western blot results have not been...

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Thank you for confirming those details.
I have a few suggestions which may help improve the results observed with ab85015.
The target you are hoping to detect is quite tricky as it is large (expected ˜226 kDa) and membrane bound. Some optimis...

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Dear,
Thank you again for the help. And indeed we've obtained quite nice results in our Flowcytometry experiments. Now however, I've tried a WB approach as a additional control experiment. I lyse the cell pellet with RIPA (+ 1mM ...

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Thank you for contacting us and updating us on the progress you have been making.
xxxxx is unfortunately away from the office at present but I will try to help as best I can on her behalf.
I'm please do hear that the antibody has been working...

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Dear,
Thank you for your information. However the inquiry more specifically concerned Immunocytochemistry, for intracellular staining of adherent cells (HEK, PC12) in a flat bottom 96-well plate. I was wondering if you have any experiences or prot...

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Thank you for your response.
We do not have specific protocol for ICC or ICC/IF for intracellular staining of adherent cells (HEK, PC12) in a flat bottom 96-well plate.
We have some general protocols under our Protocol site which could be alt...

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Dear,
Again thanks for your information. As previously asked, is it possible to use this protocol for FCM staining of cells with Nav1.7 antibody also applicable for cells that are adherent, meaning that they will be used for microscopy (ICC)? I Wa...

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Thank you for your response.
My colleague is out of office this week and she has asked me to look after her customers.
I have attached a protocol used for testing this particular antibody (ab85015). In our Lab, we have not tested it by oursel...

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Dear,

Please allow me to ask these last questions (I hope :-))

- If you're talking about centrifuging at 2.000 rpmi, it is somewhat confusing, 'cause the rotor diameter is crucial, so could you indicate then rather the ex...

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Thank you for your reply and further questions which have been forwarded to me as my colleague Karin is currently away from the office.

In answer to your questions regarding the protocol:

1. Centrifuging for Cell Fixation is 500 x...

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Dear,
Thank you very much for the info. We will repeat our staining, using your prescribed protocol. Is there no tween anymore in your washing buffer (eg 0.2% tween 20 in 1xPBS)? And do you think this protocol can be used for immunstaining og adhe...

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Thank you for getting back to me.
The wash buffer used following isolation of the cells and before fixation was just PBS. Following fixation and permeabilization, 1x PBS/1% Foetal Bovine serum was used to was the cells. I see no reason why this pr...

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1-10 of 12 Abreviews or Q&A

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