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

概述

  • 产品名称
    Anti-alpha 1 Sodium Potassium ATPase抗体[M8-P1-A3]
    参阅全部 alpha 1 Sodium Potassium ATPase 一抗
  • 描述
    小鼠单克隆抗体[M8-P1-A3] to alpha 1 Sodium Potassium ATPase
  • 宿主
    Mouse
  • 经测试应用
    适用于: IHC-Fr, WB, IHC-P, Inhibition Assay, ICC, ELISA, IP, ICC/IF, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Sheep, Chicken, Dog, Human, Pig, Drosophila melanogaster, Non human primates
  • 免疫原

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

  • 表位
    This antibody recognizes an epitope between amino acid residues 496-506 of lamb kidney sodium/potassium ATPase.
  • 阳性对照
    • WB: canine kidney extract

性能

  • 形式
    Liquid
  • 存放说明
    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.
  • 存储溶液
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • 纯度
    Protein A purified
  • 克隆
    单克隆
  • 克隆编号
    M8-P1-A3
  • 同种型
    IgG1
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab2872 in the following tested applications.

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

应用 Ab评论 说明
IHC-Fr 1/100.
WB 1/250. Detects a band of approximately 100 kDa (predicted molecular weight: 110 kDa).
IHC-P 1/100 - 1/2000.
Inhibition Assay Use at an assay dependent concentration.
ICC 1/50 - 1/200.
ELISA 1/40.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt 1/20 - 1/100.

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

 

靶标

  • 功能
    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.
  • 序列相似性
    Belongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIC subfamily.
  • 翻译后修饰
    Phosphorylation on Tyr-10 modulates pumping activity.
  • 细胞定位
    Cell membrane. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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

图片

  • 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.
  • 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.

  • 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

  • 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 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 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 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.

文献

This product has been referenced in:
See all 18 Publications for this product

客户评价及客户问答

1-6 of 6 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

提交于 Nov 04 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

提交于 Apr 03 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
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%

Ms. Amy Weaver

Verified customer

提交于 Dec 12 2005

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!

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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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