Anti-Calcium Pump PMCA4 ATPase 抗体 [JA9] (ab2783)
Key features and details
- Mouse monoclonal [JA9] to Calcium Pump PMCA4 ATPase
- Suitable for: ICC, Flow Cyt, WB, IHC-P
- Knockout validated
- Reacts with: Cow, Human
- Isotype: IgG1
製品の概要
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製品名
Anti-Calcium Pump PMCA4 ATPase antibody [JA9]
Calcium Pump PMCA4 ATPase 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [JA9] to Calcium Pump PMCA4 ATPase -
由来種
Mouse -
特異性
This antibody is specific to the PMCA4 isoform. -
アプリケーション
適用あり: ICC, Flow Cyt, WB, IHC-Pmore details -
種交差性
交差種: Cow, Human -
免疫原
Full length native protein (purified) corresponding to Human Calcium Pump PMCA4 ATPase. Purified human erythrocyte PMCA.
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ポジティブ・コントロール
- WB: SH-SY5Y and HeLa cell lysates.
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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
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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...
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精製度
Affinity purified -
ポリ/モノ
モノクローナル -
クローン名
JA9 -
アイソタイプ
IgG1 -
研究分野
関連製品
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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ab2783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 |
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ICC |
Use at an assay dependent concentration.
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Flow Cyt |
Use 2µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/1000. By Western blot, this antibody detects an ~129 kDa and an ~133 kDa protein representing PMCA4b and PMCA4a ATPase, respectively.
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IHC-P |
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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特記事項 |
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ICC
Use at an assay dependent concentration. |
Flow Cyt
Use 2µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
WB
1/1000. By Western blot, this antibody detects an ~129 kDa and an ~133 kDa protein representing PMCA4b and PMCA4a ATPase, respectively. |
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ターゲット情報
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機能
This magnesium-dependent enzyme catalyzes the hydrolysis of ATP coupled with the transport of calcium out of the cell. -
組織特異性
Isoform XB is the most abundant isoform and is expressed ubiquitously. Isoforms containing segment Z have only been detected in heart, while isoforms containing segment a have been found in heart, stomach and brain cortex. -
配列類似性
Belongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIB subfamily. -
細胞内局在
Cell membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 282146 Cow
- Entrez Gene: 493 Human
- Omim: 108732 Human
- SwissProt: P23634 Human
- Unigene: 343522 Human
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別名
- AT2B4_HUMAN antibody
- ATP2B2 antibody
- Atp2b4 antibody
see all
画像
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All lanes : Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ATP2B4 knockout HeLa cell lysate
Lane 3 : SH-SY5Y cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 140 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab2783 observed at 140 kDa. Red - loading control, ab181602 observed at 37 kDa.
ab2783 Anti-Calcium Pump PMCA4 ATPase antibody [JA9] was shown to specifically react with Calcium Pump PMCA4 ATPase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264720 (knockout cell lysate ab258321) was used. Wild-type and Calcium Pump PMCA4 ATPase knockout samples were subjected to SDS-PAGE. ab2783 and Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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IHC image of ab2783 staining in human cervical carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab2783, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Overlay histogram showing HeLa cells stained with ab2783 (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 (ab2783, 2μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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 normal biopsies of deparaffinized Human brain tissue. 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 PMCA4 ATPase ab2783 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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Immunolocalization of Calcium Pump PMCA4 ATPase in human corneal epithelium.
(A) Immunolocalization of PMCA4 (green) using ab2783 on cell membranes of all cells in each layer of human corneal epithelium.
(B) Immunolocalization of Calcium Sensing Receptor (red) using ab19347 on cell membranes of human corneal epithelium.
(C) Merged image.
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Immunolocalization of Calcium Pump PMCA4 and Calcium Sensing Receptor in bovine corneal epithelium (bCE).
(A) Immunolocalization of Calcium Pump PMCA4 (green) using ab2783 on cell membranes of all cells in each layer of bCE.
(B) Immunolocalization of Calcium Sensing Receptor (red) using ab19347 on cell membranes of bCE mostly in wing- and squamous cell layers.
(C) Merged image.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (22)
ab2783 は 22 報の論文で使用されています。
- Sun R et al. PMCA4 gene expression is regulated by the androgen receptor in the mouse testis during spermatogenesis. Mol Med Rep 23:N/A (2021). PubMed: 33355366
- Ilic K et al. Plasma Membrane Calcium ATPase-Neuroplastin Complexes Are Selectively Stabilized in GM1-Containing Lipid Rafts. Int J Mol Sci 22:N/A (2021). PubMed: 34948386
- Benedetti R et al. Inhibition of Histone Demethylases LSD1 and UTX Regulates ERa Signaling in Breast Cancer. Cancers (Basel) 11:N/A (2019). PubMed: 31888209
- Antoniou A et al. The dynamic recruitment of TRBP to neuronal membranes mediates dendritogenesis during development. EMBO Rep 19:N/A (2018). WB ; Rat . PubMed: 29263199
- Lewis S et al. Acute inhibition of PMCA4, but not global ablation, reduces blood pressure and arterial contractility via a nNOS-dependent mechanism. J Cell Mol Med 22:861-872 (2018). PubMed: 29193716