Anti-Calcineurin A antibody [EP1669Y] (ab52761)
Key features and details
- Rabbit monoclonal [EP1669Y] to Calcineurin A
- Suitable for: WB, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-Calcineurin A antibody [EP1669Y]
See all Calcineurin A primary antibodies -
Description
Rabbit monoclonal [EP1669Y] to Calcineurin A -
Host species
Rabbit -
Specificity
This antibody reacts with Calcineurin A. -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt HumanWB Human -
Immunogen
Synthetic peptide within Human Calcineurin A (N terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa, SH-SY5Y and HAP1 cell lysates. Flow Cyt: HeLa cells.
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General notes
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
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...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EP1669Y -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-Calcineurin A antibody [EP1669Y] (ab52761) at 1/1000 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 kDaLanes 1- 2: Merged signal (red and green). Green - ab52761 observed at 59 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab52761 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. ab52761 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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.
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Overlay histogram showing HeLa cells stained with ab52761 (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 (ab52761, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Calcineurin A knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: SH-SY5Y cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab52761 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab52761 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. ab52761 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.
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Anti-Calcineurin A antibody [EP1669Y] (ab52761) at 1/10000 dilution + Hela cell lysate at 10 µg
Secondary
Goat anti-rabbit HRP labeled at 1/2000 dilution
Predicted band size: 59 kDa
Observed band size: 59 kDa