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  1. Link

    calcineurin-a-antibody-epr16702-ab109412.pdf

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Immunology Adaptive Immunity B Cells Non-CD
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)

  • Datasheet
  • SDS
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Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
  • Immunocytochemistry/ Immunofluorescence - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
  • Flow Cytometry (Intracellular) - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
  • Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
  • Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
  • Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
  • Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1670(2)] to Calcineurin A
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

You may also be interested in

Protein
Product image
Recombinant Human Calcineurin A protein (ab101870)
Secondary
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Array
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Human Angiogenesis Antibody Array - Membrane (43 Targets) (ab193655)

View more associated products

Overview

  • Product name

    Anti-Calcineurin A antibody [EPR1670(2)]
    See all Calcineurin A primary antibodies
  • Description

    Rabbit monoclonal [EPR1670(2)] to Calcineurin A
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, ICC/IFmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Fetal brain, SH-SY5Y, A431, and HeLa cell lysates; HeLa cells. Mouse and rat brain cortex lysates.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR1670(2)
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • B Cells
    • Non-CD
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Phosphatases
    • Signal Transduction
    • Signaling Pathway
    • Calcium Signaling
    • Calmodulin Pathway

Associated products

  • Alternative Versions

    • Anti-Calcineurin A antibody [EPR1670(2)] - BSA and Azide free (ab239975)
    • Alexa Fluor® 647 Anti-Calcineurin A antibody [EPR1670(2)] (ab252107)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human PPP3CA (Calcineurin A) knockout HeLa cell line (ab265130)
  • KO cell lysates

    • Human PPP3CA (Calcineurin A) knockout HeLa cell lysate (ab257181)
  • Recombinant Protein

    • Recombinant Human Calcineurin A protein (ab101870)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab109412 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 (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/10000 - 1/50000. Detects a band of approximately 58 kDa (predicted molecular weight: 59 kDa).
ICC/IF
1/100 - 1/250.
Notes
Flow Cyt (Intra)
1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB
1/10000 - 1/50000. Detects a band of approximately 58 kDa (predicted molecular weight: 59 kDa).
ICC/IF
1/100 - 1/250.
Application notes
Is unsuitable for IHC-P or IP.

Target

  • Function

    Calcium-dependent, calmodulin-stimulated protein phosphatase. This subunit may have a role in the calmodulin activation of calcineurin. Dephosphorylates DNM1L, HSPB1 and SSH1.
  • Sequence similarities

    Belongs to the PPP phosphatase family. PP-2B subfamily.
  • Cellular localization

    Nucleus. Colocalizes with ACTN1 and MYOZ2 at the Z line in heart and skeletal muscle.
  • Target information above from: UniProt accession Q08209 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5530 Human
    • Entrez Gene: 19055 Mouse
    • Entrez Gene: 24674 Rat
    • Omim: 114105 Human
    • SwissProt: Q08209 Human
    • SwissProt: P63328 Mouse
    • SwissProt: P63329 Rat
    • Unigene: 435512 Human
    • Unigene: 331389 Mouse
    • Unigene: 6866 Rat
    see all
  • Alternative names

    • Alpha isoform formerly PPP2B antibody
    • Calcineurin A alpha antibody
    • Calcineurin A1 antibody
    • CalcineurinA antibody
    • Calmodulin dependent calcineurin A subunit alpha isoform antibody
    • Calmodulin-dependent calcineurin A subunit alpha isoform antibody
    • CALN antibody
    • CALNA 1 antibody
    • CALNA antibody
    • CALNA1 antibody
    • CAM PRP catalytic subunit antibody
    • CAM-PRP catalytic subunit antibody
    • CCN 1 antibody
    • CCN1 antibody
    • CNA 1 antibody
    • CNA alpha antibody
    • CNA antibody
    • CNA1 antibody
    • PP2BA_HUMAN antibody
    • PPP2B antibody
    • Ppp3ca antibody
    • Protein phosphatase 2B catalytic subunit antibody
    • Protein phosphatase 3 (formerly 2B) catalytic subunit alpha isoform antibody
    • Protein phosphatase 3 catalytic subunit alpha isoform PPP3CA antibody
    • Protein phosphatase 3 catalytic subunit alpha isozyme antibody
    • Serine/threonine protein phosphatase 2B catalytic subunit alpha isoform antibody
    • Serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform antibody
    see all

Images

  • Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    All lanes : Anti-Calcineurin A antibody [EPR1670(2)] (ab109412) at 1/10000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PPP3CA knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 59 kDa
    Observed band size: 59 kDa



    Lanes 1- 2: Merged signal (red and green). Green - ab109412 observed at 59 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab109412 was shown to react with Calcineurin A in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265130 (knockout cell lysate ab257181) was used. Wild-type HeLa and PPP3CA knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109412 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Immunocytochemistry/ Immunofluorescence - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Immunofluorescent staining of HeLa cells using 1/100 ab109412.
  • Flow Cytometry (Intracellular) - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Flow Cytometry (Intracellular) - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)

    Overlay histogram showing HeLa cells stained with ab109412 (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 (ab109412, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

     

  • Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    All lanes : Anti-Calcineurin A antibody [EPR1670(2)] (ab109412) at 1/3000 dilution

    Lane 1 : Mouse brain cortex lysate
    Lane 2 : Mouse lung lysate
    Lane 3 : Rat brain cortex lysate
    Lane 4 : Rat liver lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size: 59 kDa
    Observed band size: 58 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Calcineurin A knockout HAP1 cell lysate (20 µg)
    Lane 3: A431 cell lysate (20 µg)
    Lane 4: HeLa cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109412 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab109412 was shown to specifically react with Calcineurin A when Calcineurin A knockout samples were used. Wild-type and Calcineurin A knockout samples were subjected to SDS-PAGE. ab109412 and ab8245 (loading control to GAPDH) were both diluted 1/10000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    All lanes : Anti-Calcineurin A antibody [EPR1670(2)] (ab109412) at 1/10000 dilution

    Lane 1 : fetal brain lysate
    Lane 2 : SH-SY5Y lysate
    Lane 3 : A431 lysate
    Lane 4 : HeLa cells lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 59 kDa
    Observed band size: 58 kDa why is the actual band size different from the predicted?

  • Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)
    Anti-Calcineurin A antibody [EPR1670(2)] (ab109412)

Protocols

  • Flow cytometry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (4)

Publishing research using ab109412? Please let us know so that we can cite the reference in this datasheet.

ab109412 has been referenced in 4 publications.

  • Moon JH  et al. Calcineurin Activation by Prion Protein Induces Neurotoxicity via Mitochondrial Reactive Oxygen Species. Oxid Med Cell Longev 2021:5572129 (2021). PubMed: 34394828
  • Gu LF  et al. Huangkui Capsule Ameliorates Renal Fibrosis in a Unilateral Ureteral Obstruction Mouse Model Through TRPC6 Dependent Signaling Pathways. Front Pharmacol 11:996 (2020). PubMed: 32719603
  • Liu Z  et al. Discovery of Small-Molecule Inhibitors of the HSP90-Calcineurin-NFAT Pathway against Glioblastoma. Cell Chem Biol 26:352-365.e7 (2019). PubMed: 30639261
  • Tian CJ & Pang X Ca2+-calcineurin signaling is involved in norepinephrine-induced cardiac fibroblasts activation. Int J Clin Exp Pathol 8:5210-6 (2015). PubMed: 26191219

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