概述

  • 产品名称Anti-ATP1A2抗体[M7-PB-E9]
    参阅全部 ATP1A2 一抗
  • 描述
    小鼠单克隆抗体[M7-PB-E9] to ATP1A2
  • 特异性Detects pan alpha sodium / potassium ATPase. The antibody binds all alpha subunits in sheep, dog, pig, human and chicken. It does not bind alpha 1 or 2 in rat, but does bind alpha 3.
  • 经测试应用适用于: WB, ICC/IF, Inhibition Assay, ICC, Flow Cyt, IHC-P, IP, ELISA, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Sheep, Chicken, Cow, Dog, Human, Pig
  • 免疫原

    Full length native protein (purified) corresponding to Sheep ATP1A2. Puified from Sheep Kidney.

  • 表位This antibody recognizes an epitope between amino acid residues 646 and 652 of the sheep kidney alpha sodium / potassium ATPase.
  • 阳性对照
    • In western blot, this antibody gave a positive signal in human, mouse and rat kidney tissue lysates.

性能

应用

Our Abpromise guarantee covers the use of ab2871 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
WB 1/500. Detects a band of approximately 102 kDa (predicted molecular weight: 114 kDa).
ICC/IF 1/20.
Inhibition Assay Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
Flow Cyt 1/100.
IHC-P Use at an assay dependent concentration.
IP 1/100 - 1/200.
ELISA Use at an assay dependent concentration.
IHC-Fr 1/100.

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, providing the energy for active transport of various nutrients.
  • 疾病相关Defects in ATP1A2 are the cause of migraine familial hemiplegic type 2 (FHM2) [MIM:602481]. FHM2 is a rare, severe, autosomal dominant subtype of migraine characterized by aura and some hemiparesis.
    Defects in ATP1A2 are a cause of alternating hemiplegia of childhood (AHC) [MIM:104290]. AHC is typically distinguished from familial hemiplegic migraine by infantile onset of the symptoms and high prevalence of associated neurological deficits that become increasingly obvious with age.
  • 序列相似性Belongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIC subfamily.
  • 细胞定位Membrane. Cell membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • AT1A2_HUMAN antibody
    • Atp1a2 antibody
    • FHM2 antibody
    • KIAA0778 antibody
    • MHP2 antibody
    • Na(+)/K(+) ATPase alpha-2 subunit antibody
    • Na+/K+ ATPase alpha 2 subunit antibody
    • Sodium potassium ATPase antibody
    • Sodium pump subunit alpha 2 antibody
    • Sodium pump subunit alpha-2 antibody
    • Sodium/potassium transporting ATPase alpha 2 chain antibody
    • Sodium/potassium transporting ATPase subunit alpha 2 antibody
    • Sodium/potassium-transporting ATPase subunit alpha-2 antibody
    see all

Anti-ATP1A2 antibody [M7-PB-E9] 图像

  • ATP1A2 was immunoprecipitated using 0.5mg Mouse Kidney tissue extract, 5µg of Mouse monoclonal to ATP1A2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Kidney tissue extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab2871.

    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 102kDa; ATP1A2

  • This image was kindly supplied as part of the review submitted by Marko Nykanen.
  • ICC/IF image of ab2871 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2871, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • All lanes : Anti-ATP1A2 antibody [M7-PB-E9] (ab2871) at 1/500 dilution

    Lane 1 : Human kidney tissue lysate - total protein (ab30203)
    Lane 2 : Kidney (Mouse) Tissue Lysate
    Lane 3 : Kidney (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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 : 114 kDa
    Observed band size : 102 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 28 kDa,60 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 2 minutes
  • Overlay histogram showing HEK293 cells stained with ab2871 (red line). The cells were fixed with 80% 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 (ab28711, 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.

    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 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 1:100 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha ab2871 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 testis 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 1:200 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha ab2871 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 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 1:200 with a mouse monoclonal antibody recognizing Sodium/Potassium ATPase alpha ab2871 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.

  • Immunocytochemistry/Immunofluorescence analysis of ATP1A2 shows staining in HeLa cells. ATP1A2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2871 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of ATP1A2 shows staining in MCF-7 cells. ATP1A2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2871 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of ATP1A2 shows staining in U251 cells. ATP1A2 (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2871 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

Anti-ATP1A2 antibody [M7-PB-E9] (ab2871)参考文献

This product has been referenced in:
  • Ait-Omar A  et al. GLUT2 Accumulation in Enterocyte Apical and Intracellular Membranes: A Study in Morbidly Obese Human Subjects and ob/ob and High Fat-Fed Mice. Diabetes 60:2598-607 (2011). WB, IHC-Fr ; Human . Read more (PubMed: 21852673) »
  • Jiang J  et al. Localization of Na+-K+ ATPases in quasi-native cell membranes. Nano Lett 9:4489-93 (2009). Human . Read more (PubMed: 19807066) »

See all 3 Publications for this product

Product Wall

Ab2871 binds all alpha subunits in sheep, dog, pig, human and chicken. It does not bind alpha 1 or 2 in rat, but does bind alpha 3. Unfortunately we do not have a sodium potassium ATPase antibody that is specific for alpha 2. If you have any ad...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"