Anti-PPP3CA Antibody Picoband™

Product Name

Anti-PPP3CA Antibody Picoband™

View all Calcineurin A Antibodies

SKU/Catalog Number

A03026-3

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-PPP3CA Antibody Picoband™ catalog # A03026-3. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-PPP3CA Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A03026-3)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human PPP3CA recombinant protein (Position: E3-D511).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A03026-3 is reactive to PPP3CA in Human, Rat

Applications

A03026-3 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

59 kDa

Calculated molecular weight

58.688kDa

Background of Calcineurin A

Calcineurin A is also known as PPP3CA. It is mapped to 4q24. Semsarian et al. (1999) and Musaro et al. (1999) independently showed that IGF1stimulates skeletal muscle hypertrophy and a switch to glycolytic metabolism by activating calcineurin A and inducing the nuclear translocation of transcription factor NFATC1. Semsarian et al. (1999) found that hypertrophy was suppressed by the calcineurin inhibitors cyclosporin A or FK506, but not by inhibitors of the MAP kinase or phosphatidylinositol-3-OH kinase pathways. Musaro et al. (1999) showed that expression of a dominant-negative calcineurin mutant also repressed myocyte differentiation and hypertrophy. Musaro et al. (1999) demonstrated that either IGF1 or activated calcineurin induces expression of transcription factor GATA2, which accumulates in a subset of myocyte nuclei, where it associates with calcineurin and a specific dephosphorylated isoform of NFATC1.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Human, Rat
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of PPP3CA using anti-PPP3CA antibody (A03026-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human U-87MG whole cell lysates,
Lane 3: rat C6 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-PPP3CA antigen affinity purified polyclonal antibody (Catalog # A03026-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for PPP3CA at approximately 59 kDa. The expected band size for PPP3CA is at 59 kDa.

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Figure 2. IF analysis of PPP3CA using anti-PPP3CA antibody (A03026-3).
PPP3CA was detected in an immunocytochemical section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-PPP3CA Antibody (A03026-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of SiHa cells using anti-PPP3CA antibody (A03026-3).
Overlay histogram showing SiHa cells stained with A03026-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PPP3CA Antibody (A03026-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For PPP3CA (Source: Uniprot.org, NCBI)

Alternative Names

calcineurin A alpha; Calcineurin A; Calmodulin-dependent calcineurin A subunit alpha isoform; CALN; CALNA; CALNAcatalytic subunit, alpha isoform; CAM-PRP catalytic subunit; CNA; CNA1; CNA1CALNA1; EC 3.1.3.16; PP2B; PP3CA; PPP2B; PPP2BCCN1; PPP3CA; protein phosphatase 2B, catalytic subunit, alpha isoform; protein phosphatase 3 (formerly 2B), catalytic subunit, alpha isoform(calcineurin A alpha); protein phosphatase 3, catalytic subunit, alpha isozyme; serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform PPP3CA ACCIID, CALN, CALNA, CALNA1, CCN1, CNA1, DEE91, IECEE, IECEE1, PPP2B protein phosphatase 3 catalytic subunit alpha serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform|CAM-PRP catalytic subunit|calcineurin A alpha|protein phosphatase 3 catalytic subunit alpha isozyme

For more info on PPP3CA, check out the PPP3CA Infographic

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In this infographic, you will see the following information for PPP3CA: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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