Key features and details
- Mouse monoclonal [JA9] to Calcium Pump PMCA4 ATPase
- Suitable for: ICC, Flow Cyt, WB, IHC-P
- Knockout validated
- Reacts with: Rat, Rabbit, Cow, Human, Pig
- Isotype: IgG1
产品名称Anti-Calcium Pump PMCA4 ATPase抗体[JA9]
参阅全部 Calcium Pump PMCA4 ATPase 一抗
描述小鼠单克隆抗体[JA9] to Calcium Pump PMCA4 ATPase
特异性This antibody is specific to the PMCA4 isoform.
经测试应用适用于: ICC, Flow Cyt, WB, IHC-Pmore details
种属反应性与反应: Rat, Rabbit, Cow, Human, Pig
Other Immunogen Type corresponding to Human Calcium Pump PMCA4 ATPase. Purified human erythrocyte PMCA.
- WB: SH-SY5Y and HeLa cell lysates.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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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...
KO cell lines
KO cell lysates
Our Abpromise guarantee covers the use of ab2783 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|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.|
功能This magnesium-dependent enzyme catalyzes the hydrolysis of ATP coupled with the transport of calcium out of the cell.
组织特异性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.
序列相似性Belongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIB subfamily.
- Information by UniProt
- AT2B4_HUMAN antibody
- ATP2B2 antibody
- Atp2b4 antibody
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.
All lanes : Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783) at 1/1000 dilution
All lanes : Rat cortical neurons whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : HRP conjugated goat anti-mouse
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 130 kDa why is the actual band size different from the predicted?
Exposure time: 2 seconds
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.
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.
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.
ab2783 被引用在 20 文献中.
- 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
- Ezra-Nevo G et al. Cerebellar Learning Properties Are Modulated by the CRF Receptor. J Neurosci 38:6751-6765 (2018). PubMed: 29934353
- Frische S et al. H+-ATPase B1 subunit localizes to thick ascending limb and distal convoluted tubule of rodent and human kidney. Am J Physiol Renal Physiol 315:F429-F444 (2018). PubMed: 29993276