Anti-pan ATPase Alpha抗体[M7-PB-E9] (ab2871)
Key features and details
- Mouse monoclonal [M7-PB-E9] to pan ATPase Alpha
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IHC-P, IP
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
概述
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产品名称
Anti-pan ATPase Alpha抗体[M7-PB-E9] -
描述
小鼠单克隆抗体[M7-PB-E9] to pan ATPase Alpha -
宿主
Mouse -
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, IHC-P, ICC/IF, Flow Cyt, IHC-P, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Full length native protein (purified) corresponding to Sheep pan ATPase Alpha. Puified from Sheep Kidney.
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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.
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常规说明
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
性能
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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
Constituent: 0.1% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
M7-PB-E9 -
同种型
IgG1 -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
应用
应用 | Ab评论 | 说明 |
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WB |
1/500. Detects a band of approximately 102 kDa (predicted molecular weight: 114 kDa).
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IHC-P |
1/100 - 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/20.
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Flow Cyt |
1/100.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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IHC-P |
Use at an assay dependent concentration.
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IP |
1/100 - 1/200.
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说明 |
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WB
1/500. Detects a band of approximately 102 kDa (predicted molecular weight: 114 kDa). |
IHC-P
1/100 - 1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/20. |
Flow Cyt
1/100. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
IHC-P
Use at an assay dependent concentration. |
IP
1/100 - 1/200. |
靶标
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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
Migraine, familial hemiplegic, 2
Alternating hemiplegia of childhood 1 -
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
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数据库链接
- Entrez Gene: 477 Human
- SwissProt: P50993 Human
- Unigene: 34114 Human
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别名
- AT1A2_HUMAN antibody
- Atp1a2 antibody
- Na(+)/K(+) ATPase alpha-2 subunit antibody
see all
图片
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All lanes : Anti-pan ATPase Alpha 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
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: 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 -
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
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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.
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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 paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)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.
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Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)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.
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Immunohistochemistry paraffin embedded sections - Anti-pan ATPase Alpha antibody [M7-PB-E9] (ab2871)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.
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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.
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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.
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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.
实验方案
数据表及文件
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Datasheet download
文献 (5)
ab2871 被引用在 5 文献中.
- Zhao C et al. ß-Catenin Controls the Electrophysiologic Properties of Skeletal Muscle Cells by Regulating the a2 Isoform of Na+/K+-ATPase. Front Neurosci 13:831 (2019). PubMed: 31440132
- Al-Abdulla R et al. Epigenetic events involved in organic cation transporter 1-dependent impaired response of hepatocellular carcinoma to sorafenib. Br J Pharmacol 176:787-800 (2019). PubMed: 30592786
- 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 . PubMed: 21852673
- Jiang J et al. Localization of Na+-K+ ATPases in quasi-native cell membranes. Nano Lett 9:4489-93 (2009). Human . PubMed: 19807066
- Ball WJ et al. Isolation and characterization of monoclonal antibodies to (Na+ + K+)-ATPase. Biochim Biophys Acta 719:413-23 (1982). PubMed: 6295504