Anti-alpha 1 Sodium Potassium ATPase 抗体 [M8-P1-A3] (ab2872)

Reviews (3)Q&A (3)References (20)
Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
  • Flow Cytometry - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
  • Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)

Key features and details

  • Mouse monoclonal [M8-P1-A3] to alpha 1 Sodium Potassium ATPase
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
  • Reacts with: Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3]
    alpha 1 Sodium Potassium ATPase 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [M8-P1-A3] to alpha 1 Sodium Potassium ATPase

  • 由来種

    Mouse

  • アプリケーション

    適用あり: WB, IHC-P, ICC/IF, Flow Cytmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Other Immunogen Type corresponding to Sheep alpha 1 Sodium Potassium ATPase. Lamb kidney alpha 1 sodium/potassium ATPase.

  • エピトープ

    This antibody recognizes an epitope between amino acid residues 496-506 of lamb kidney sodium/potassium ATPase.

  • ポジティブ･コントロール

    • WB: canine kidney extract

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    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.

  • バッファー

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...

  • 精製度

    Protein A purified

  • ポリ/モノ

    モノクローナル

  • クローン名

    M8-P1-A3

  • アイソタイプ

    IgG1

  • 研究分野

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab2872の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (1)
1/200 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 110 kDa).
IHC-P (2)
1/100 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50 - 1/200.
Flow Cyt
1/20 - 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
特記事項
WB
1/200 - 1/2000. Detects a band of approximately 100 kDa (predicted molecular weight: 110 kDa).
IHC-P
1/100 - 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/50 - 1/200.
Flow Cyt
1/20 - 1/100.

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

ターゲット情報

  • 機能

    This is the catalytic component of the active enzyme, which catalyzes the hydrolysis of ATP coupled with the exchange of sodium and potassium ions across the plasma membrane. This action creates the electrochemical gradient of sodium and potassium ions, providing the energy for active transport of various nutrients.

  • 配列類似性

    Belongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIC subfamily.

  • 翻訳後修飾

    Phosphorylation on Tyr-10 modulates pumping activity.

  • 細胞内局在

    Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt

  • 参照データベース

    • 別名

      • A1A1 antibody
      • AT1A1 antibody
      • AT1A1_HUMAN antibody
      • ATP1A1 antibody
      • Atpa-1 antibody
      • ATPase Na+/K+ transporting alpha 1 polypeptide antibody
      • ATPase Na+/K+ transporting subunit alpha 1 antibody
      • BC010319 antibody
      • EC 3.6.3.9 antibody
      • MGC3285 antibody
      • MGC38419 antibody
      • MGC51750 antibody
      • Na K ATPase alpha A catalytic polypeptide antibody
      • Na K ATPase catalytic subunit alpha A protein antibody
      • Na(+)/K(+) ATPase 1 antibody
      • Na(+)/K(+) ATPase alpha-1 subunit antibody
      • Na+, K+ ATPase alpha subunit antibody
      • Na+/K+ ATPase alpha 1 subunit antibody
      • Na+/K+ ATPase 1 antibody
      • Na,K ATPase alpha 1 subunit antibody
      • Nkaa1b antibody
      • Sodium potassium ATPase alpha 1 polypeptide antibody
      • Sodium pump 1 antibody
      • Sodium pump subunit alpha-1 antibody
      • sodium-potassium ATPase catalytic subunit alpha-1 antibody
      • Sodium/potassium-transporting ATPase subunit alpha-1 antibody
      see all

    画像

    • Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      All lanes : Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872) at 1/1000 dilution

      Lane 1 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
      Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
      Lane 3 : MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/4000 dilution

      Predicted band size: 110 kDa



      Samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel. Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System.

      Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit.

    • Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunocytochemistry/ Immunofluorescence - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)

      Immunocytochemistry/Immunofluorescence analysis of alpha 1 Sodium Potassium ATPase (green) in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes at room temperature and blocked with 0.3% BSA for 15 minutes at room temperature. Cells were incubated with ab2872 (1:100) for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (1:500) for 30 minutes at room temperature. F-actin (red) was stained with DyLight 594 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)This image is courtesy of an anonymous Abreview

      ab2872 staining pig hepatocyte tissue sections by IHC-P.  The section was fixed with Bouins and subjected to heat mediated antigen retrieval (at pH 9) prior to incubating with the primary antibody, diluted 1/2000, for 1 hour at 20°C.  A Cy3® conjugated goat anti-mouse IgG antibody was used as the secondary.

      See Abreview

    • Flow Cytometry - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Flow Cytometry - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Overlay histogram showing HEK293 cells stained with ab2872 (red line). The cells were fixed with 100% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2872, 1/100 dilution) 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 signal in HEK293 cells fixed with 4% paraformaldehyde used under the same conditions.

      Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:50 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha-1 ab2872 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human kidney tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha-1 ab2872 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha-1 ab2872 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

    • Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      Western blot - Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872)
      All lanes : Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3] (ab2872) at 1/500 dilution

      Lane 1 : Human brain normal tissue lysate - membrane extract (ab29456)
      Lane 2 : Human testis tissue lysate - total protein (ab30257)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 110 kDa
      Observed band size: 100 kDa why is the actual band size different from the predicted?
      Additional bands at: 25 kDa, 70 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 8 minutes


      The 100 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to alpha 1 Sodium Potassium ATPase.

    参考文献 (20)

    ab2872 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。

    ab2872 は 20 報の論文で使用されています。

    • Zhang SH  et al. A CASPR1-ATP1B3 protein interaction modulates plasma membrane localization of Na+/K+-ATPase in brain microvascular endothelial cells. J Biol Chem 294:6375-6386 (2019). PubMed: 30792309
    • Zhang F  et al. Molecular mechanisms of caffeine-mediated intestinal epithelial ion transports. Br J Pharmacol 176:1700-1716 (2019). PubMed: 30808064
    • Zhang D  et al. Downregulation of ATP1A1 promotes cancer development in renal cell carcinoma. Clin Proteomics 14:15 (2017). WB, IHC ; Human . PubMed: 28484360
    • Zhu W  et al. Effect of docosahexaenoic acid on traumatic brain injury in rats. Exp Ther Med 14:4411-4416 (2017). PubMed: 29075341
    • Stordal B  et al. Resistance to paclitaxel in a cisplatin-resistant ovarian cancer cell line is mediated by P-glycoprotein. PLoS One 7:e40717 (2012). WB ; Human . PubMed: 22792399
    View all Publications for this product

    レビューと Q&A

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    1-6 of 6 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Hepatocytes)
    Specification
    Hepatocytes
    Fixative
    Bouins
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 9,0
    Permeabilization
    No
    Read More

    Abcam user community

    Verified customer

    投稿 Nov 04 2008

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3]

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (Hepatocytes)
    Specification
    Hepatocytes
    Fixative
    Bouins
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: pH 9,0
    Permeabilization
    No
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 03 2008

    Western blot abreview for Anti-alpha 1 Sodium Potassium ATPase antibody [M8-P1-A3]

     Excellent
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Glioma)
    Specification
    Glioma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Ms. Amy Weaver

    Verified customer

    投稿 Dec 12 2005

    Question

    Thank you very much for kindly answering my questions and i would like to know how long does it takes to have ab2872 antibody with new lot number. I would like to have this antibody with new lot number.

    Read More
    Answer

    Thank you for your reply. We are going to send you a new batch as soon as we receive it. This is to let you know that I have placed a new order for you - one vial of ab2872 free of charge. For your information your new order number is: 83747. Thank you for your patience!

    Read More

    Question

    1.actually the secondary antibody which i am using is anti-mouse IgG, Horseradish Peroxidase linked whole antibody(from sheep) from amersham. 2. i have been doing western blots from past 5 years. 3.i am following the same protocol for all other antibodies even your other abcam antibodies, i never experienced this problem. 4.As per my knowledge the incubation time doesn't matter either overnight or 2 hours. 5. i don't think reducing the boiling time will make difference. could you please replace with the new antibody.

    Read More
    Answer

    Thank you for getting back to me and providing more information about your Western blot protocol. We can offer you a new vial as a free of charge replacement; however it is from the exatly the same batch. Unfortunately, we do not have different batch in our stock at the moment. We would like to suggest loading less protein than 45 ug per lane, the optimal loading should be 20-30 ug per lane. We do understand and appreciate your experience doing Western blot assay. Using standard protocol is useful; however each target protein and antibody needs optimization. Boiling can make a difference and if the samples are over-boiled, it can change the detection of the target protein. Try to boil the samples for 3 or max 5 min. Please so let me know if you would like to get the vial.

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    Question

    BATCH NUMBER 90665 ORDER NUMBER 66788 DESCRIPTION OF THE PROBLEM Multiple bands SAMPLE rat np cell lysates and we couldn't see bands in rat tissues like kidney, cartilage, liver and heart PRIMARY ANTIBODY 1-250 dilution as recommended in protocol of ab2872. incubated for O/N at 4 degrees. 3 times wash with 1X TBST - 10 min/wash DETECTION METHOD ECL advanced POSITIVE AND NEGATIVE CONTROLS USED I used for rat kidney tissue ANTIBODY STORAGE CONDITIONS -20 SAMPLE PREPARATION i lysis the cells from MPER buffer and i used protease cocktail. I heated the samples with 5x sample buffer for 7-10 min. AMOUNT OF PROTEIN LOADED 45ug ELECTROPHORESIS/GEL CONDITIONS SDS page gel TRANSFER AND BLOCKING CONDITIONS Transfer - ready to use buffer from biorad, 70 volts for 2 hours at 4 degrees. 5% non fat dry milk in 1X TBST SECONDARY ANTIBODY 1-2000 dilution with HRP mouse monoclonal from amersham, incubated at RT for 1 1/2 hour. 3 times wash with 1X TBST - 10 min/wash HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? first time i used rat tissue lysates, i see multiple bands and then i tried with rat NP cell lysates. I got multiple bands again ADDITIONAL NOTES If u want i can send the blot which i ran. i never got these multiple bands with ur antibodies which i used before like HSP-70(very clean and good antibody). i am getting the bands only with this particular antibody. I would like request you to replace this antibody with new batch of same antibody

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    Answer

    Thank you for contacting us regarding ab2872 and for taking your time to fill in the on-line Questionnaire. We are very sorry to hear that you are experiencing problem with this antibody. As we understand from your e-mail, you have used HRP mouse monoclonal as secondary antibody. Ab2872 is Mouse monoclonal antibody to alpha 1 Sodium Potassium ATPase so the compatible secondary antibody should be raised in a different species rather than mouse. Could you please specify again in which species the secondary antibody was raised? We would suggest loading 25-30 ug protein per lane, 45 ug is probably a bit too much. Try to boil the samples for 3-5 min but not longer than 5 min and incubate the membrane for 1 or 2 hrs instead of overnight. Should you still have problem with this antibody, then please do not hesitate to contact us again.

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