Validated using a knockout cell line

Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)

Reviews (1)Q&A (1)References (20)
Western blot - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
  • Flow Cytometry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
  • Immunocytochemistry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
  • Immunocytochemistry - Anti-Calcium Pump PMCA4 ATPase antibody [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

Overview

  • Product name

    Anti-Calcium Pump PMCA4 ATPase antibody [JA9]
    See all Calcium Pump PMCA4 ATPase primary antibodies

  • Description

    Mouse monoclonal [JA9] to Calcium Pump PMCA4 ATPase

  • Host species

    Mouse

  • Specificity

    This antibody is specific to the PMCA4 isoform.

  • Tested applications

    Suitable for: ICC, Flow Cyt, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Cow, Human

  • Immunogen

    Other Immunogen Type corresponding to Human Calcium Pump PMCA4 ATPase. Purified human erythrocyte PMCA.

  • Positive control

    • WB: SH-SY5Y and HeLa cell lysates.

  • General notes

    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

Properties

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
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)
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.
Notes
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.

Target

  • Function

    This magnesium-dependent enzyme catalyzes the hydrolysis of ATP coupled with the transport of calcium out of the cell.

  • Tissue specificity

    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.

  • Sequence similarities

    Belongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIB subfamily.

  • Cellular localization

    Cell membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • AT2B4_HUMAN antibody
      • ATP2B2 antibody
      • Atp2b4 antibody
      • ATPase Ca++ transporting plasma membrane 4 antibody
      • Cation transporting ATPase antibody
      • DKFZp686G08106 antibody
      • DKFZp686M088 antibody
      • Matrix remodelling associated 1 antibody
      • Matrix remodelling associated protein 1 antibody
      • Matrix-remodeling-associated protein 1 antibody
      • MXRA 1 antibody
      • MXRA1 antibody
      • OTTHUMP00000034088 antibody
      • OTTHUMP00000034089 antibody
      • Plasma membrane calcium ATPase 4 antibody
      • Plasma membrane calcium ATPase isoform 4 antibody
      • Plasma membrane calcium pump isoform 4 antibody
      • Plasma membrane calcium transporting ATPase 4 antibody
      • Plasma membrane calcium-transporting ATPase 4 antibody
      • PMCA 4 antibody
      • PMCA4 antibody
      • PMCA4b antibody
      • PMCA4x antibody
      • Sarcolemmal calcium pump antibody
      see all

    Images

    • Western blot - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      Western blot - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      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.

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      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.

    • Flow Cytometry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      Flow Cytometry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)

      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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      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.

    • Immunocytochemistry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      Immunocytochemistry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)

      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.

    • Immunocytochemistry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)
      Immunocytochemistry - Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783)

      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.

    References (20)

    Publishing research using ab2783? Please let us know so that we can cite the reference in this datasheet.

    ab2783 has been referenced in 20 publications.

    • 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
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Question

    I would like ask some questions about your products: Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783) or Anti-Calcium Pump pan PMCA ATPase antibody [5F10] (ab2825). We are making experiment on activity of Ca2+-ATPase (PMCA), so we have information about functionality and now we would like to make some new experiments about amount of this PMCA (quantity) and searching some possibilities how to do this. I'm new in this subject so excusive me if my question will be beginner question. I found it is possible to measure this with flow cytometry, it was told me that it is necessarily for flow cytometry measurements that the antibodies must be fluorescein-labeled antibodies. I didn't see anything about this in your product description, although the aplication is flow cytometry. I would like to ask if this antibodies are fluorescein-labeled? If not how is possible to make flow cytometry measurements? Second question I have: In product informations you show results on Jurkat cells, but our sample is human erythrocytes. It is possible to use this antibody on RBC?What's the conditions? In the second antibody (ab2825) it is written that: "Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Cat, Dog, Human, Syrian Hamster" and than "Predicted to work with Non Human Primates". It is suitable to use it in human? Thank you very much for your time and willingness.

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    Answer

    Thank you for contacting us.

    ab2825 can be used in flow cytometry and is positively tested however for detection you would need a Fluorophore conjugated anti mouse secondary antibody. Please click the below link for protocol;

    https://www.abcam.com/index.html?pageconfig=resource&rid=11381

    The compatible secondary antibodies are

    https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-heavy-chain-FITC-ab97239.html

    https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG1-heavy-chain-FITC-pre-adsorbed-ab98692.html

    https://www.abcam.com/Rat-monoclonal-SB77e-Secondary-Antibody-to-Mouse-IgG1-FITC-pre-adsorbed-ab133859.html

    https://www.abcam.com/Mouse-IgG-secondary-antibody-LO-MG-1-2-IgG1-Heavy-Chain-specific-ab11588.html

    We have tested ab2783 with Hela cells and is fully guaranteed for all Rat and human cells require they express the protein. Human erythrocytes can be used with this antibody.

    ab2825 has been tested with human Jurkat and monocytic cells so it will work with all PMCA4 expressing human cells.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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    Western blot abreview for Anti-Calcium Pump PMCA4 ATPase antibody [JA9]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (primary cortical neurons)
    Loading amount
    20 µg
    Specification
    primary cortical neurons
    Gel Running Conditions
    Reduced Denaturing
    Read More

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    Submitted Jan 17 2008

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