Overview

  • Product nameAnti-NCX1 antibody [C2C12]
    See all NCX1 primary antibodies
  • Description
    Mouse monoclonal [C2C12] to NCX1
  • Tested applicationsSuitable for: IHC-P, ICC/IF, IHC-Fr, ELISA, IP, WB, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Guinea pig, Dog, Human, Pig
  • Immunogen

    Full length native protein (purified) corresponding to Dog NCX1. Purified from canine cardiac sodium/calcium exchanger.

  • EpitopeThis antibody recognizes an epitope between amino acids 371-525 on the intracellular side of the plasma membrane.

Properties

Applications

Our Abpromise guarantee covers the use of ab2869 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
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/200.
IHC-Fr Use at an assay dependent concentration. PubMed: 21408028
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB 1/1000. By Western blot, this antibody detects a 120 kDa protein representing the sodium / calcium exchanger from guinea pig cardiac extract. The bands seen at 70 kDa and 160 kDa represent a proteolytic fragment and non-reduced exchanger respectively. This antibody is not recommended for Western blot procedures of rat tissues.
Flow Cyt 1/20 - 1/100.

Target

Anti-NCX1 antibody [C2C12] images

  • Immunocytochemistry/Immunofluorescence analysis of NCX1 shows staining in A2058 cells. NCX1 staining (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 ab2869 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of NCX1 shows staining in A549 cells. NCX1 staining (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 ab2869 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of NCX1 shows staining in U251 cells. NCX1 staining (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 ab2869 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • ab2869 staining NCX1 in Human kidney tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde, permeabilized with 0.05% Tween20 and blocked with 5% normal goat serum in 1XPBS + 0.05% Tween20 for 1 hour at 25°C; antigen retrieval was by heat mediation in sodium citrate (pH 6.0) buffer. Samples were incubated with primary antibody (1/100 in blocking buffer) for 1 hour at 25°C. Ab47827 (1/500) was used as the secondary antibody.

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  • This image shows Human embryonic stem cell derived cardiomyocytes, stained with dapi (blue) and anti-NCX1 antibody ab2869 (red). The cells were fixed in paraformaldehyde, permeabilized with 0.1% Triton X-100 in PBS and blocked with 4% goat serum for 1 hour. The cells were then incubated with primary antibody (1/100) for 16 hours at 4°C.

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  • Overlay histogram showing HEK293 cells stained with ab2869 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2869, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgM (mu chain) (ab97007) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • Immunohistochemistry was performed on normal deparaffinized Human kidney 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 1:50 with a mouse monoclonal antibody recognizing Sodium/Calcium Exchanger ab2869 or without primary antibody (negative control; right panel) 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.

  • Immunohistochemistry was performed on normal deparaffinized Human heart 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 1:20 with a mouse monoclonal antibody recognizing Sodium/Calcium Exchanger ab2869 or without primary antibody (negative control; right panel) 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.

References for Anti-NCX1 antibody [C2C12] (ab2869)

This product has been referenced in:
  • Hong H  et al. Elevated NCX1 and NCKX4 expression in the patent postnatal ductus arteriosus of ductal-dependent congenital heart disease patients. Pediatr Cardiol 36:743-51 (2015). Read more (PubMed: 25500693) »
  • Raynaud CM  et al. Human embryonic stem cell derived mesenchymal progenitors express cardiac markers but do not form contractile cardiomyocytes. PLoS One 8:e54524 (2013). ICC/IF ; Human . Read more (PubMed: 23342164) »

See all 6 Publications for this product

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (hESC-derived cardiomyocyte)
Specification hESC-derived cardiomyocyte
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton X-100 in PBS
Blocking step Goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
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Submitted Nov 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Normal Kidney)
Specification Normal Kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium Citrate pH6.0
Permeabilization Yes - 0.05% Tween20
Blocking step Normal Goat Serum in 1X PBS (0.05% Tween20) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 20 2010

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