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

Overview

  • Product nameAnti-Calcium Pump PMCA4 ATPase antibody [JA9]
    See all Calcium Pump PMCA4 ATPase primary antibodies
  • Description
    Mouse monoclonal [JA9] to Calcium Pump PMCA4 ATPase
  • SpecificityThis antibody is specific to the PMCA4 isoform.
  • Tested applicationsSuitable for: Flow Cyt, WB, IHC-P, ELISA, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Cow, Human, Pig
  • Immunogen

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

Properties

Applications

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
Flow Cyt Use 2µg for 106 cells.
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.
ELISA 1/15000.
IHC-Fr 1/100 - 1/500.
ICC/IF Use at an assay dependent concentration. PubMed: 18229969

Target

  • FunctionThis magnesium-dependent enzyme catalyzes the hydrolysis of ATP coupled with the transport of calcium out of the cell.
  • Tissue specificityIsoform 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 similaritiesBelongs to the cation transport ATPase (P-type) (TC 3.A.3) family. Type IIB subfamily.
  • Cellular localizationCell 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

Anti-Calcium Pump PMCA4 ATPase antibody [JA9] images

  • All lanes : Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783) at 1/1000 dilution

    Lane 1 : Rat cortical neurons whole cell lysate
    Lane 2 : Rat cortical neurons whole cell lysate
    Lane 3 : Rat cortical neurons whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    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

    This image is courtesy of an anonymous Abreview

    See Abreview

  • 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.

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

This product has been referenced in:
  • Kosiorek M  et al. Interaction of plasma membrane Ca(2+)-ATPase isoform 4 with calcineurin A: Implications for catecholamine secretion by PC12 cells. Biochem Biophys Res Commun 411:235-40 (2011). WB ; Rat . Read more (PubMed: 21740891) »
  • Mohamed TM  et al. Plasma membrane calcium pump (PMCA4)/neuronal nitric oxide synthase complex regulates cardiac contractility through modulation of a compartmentalized cyclic nucleotide microdomain. J Biol Chem : (2011). WB ; Mouse . Read more (PubMed: 21965681) »

See all 8 Publications for this product

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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;

http://w...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
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Verified customer

Submitted Jan 17 2008

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