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AB257181

Human PPP3CA (Calcineurin A) knockout HeLa cell lysate

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PPP3CA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1.
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Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell lysate (AB257181)
  • WB

Lab

Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell lysate (AB257181)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : PPP3CA knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab52761 observed at 59 kDa. Red - loading control ab8245 observed at 37 kDa.

ab52761 Rabbit monoclonal [EP1669Y] to Calcineurin A was shown to specifically 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 and Calcineurin A knockout samples 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) were incubated overnight at 4°C at 1 in 1000 dilution and 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.

All lanes:

Western blot - Anti-Calcineurin A antibody [EP1669Y] (<a href='/en-us/products/primary-antibodies/calcineurin-a-antibody-ep1669y-ab52761'>ab52761</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PPP3CA knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ppp3ca-calcineurin-a-knockout-hela-cell-line-ab265130'>ab265130</a>)

Predicted band size: 59 kDa

Observed band size: 59 kDa

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Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell lysate (AB257181)
  • WB

Lab

Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell lysate (AB257181)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : PPP3CA knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab109412 observed at 59 kDa. Red - loading control ab8245 observed at 37 kDa.

ab109412 Rabbit monoclonal [EPR1670(2)] to Calcineurin A was shown to specifically 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 and Calcineurin A knockout samples 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) were incubated overnight at 4°C at 1 in 10000 dilution and 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.

All lanes:

Western blot - Anti-Calcineurin A antibody [EPR1670(2)] (<a href='/en-us/products/primary-antibodies/calcineurin-a-antibody-epr16702-ab109412'>ab109412</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PPP3CA knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ppp3ca-calcineurin-a-knockout-hela-cell-line-ab265130'>ab265130</a>)

Predicted band size: 59 kDa

Observed band size: 59 kDa

false

Sanger Sequencing - Human PPP3CA (Calcineurin A) knockout HeLa cell lysate (AB257181)
  • Sanger seq

Unknown

Sanger Sequencing - Human PPP3CA (Calcineurin A) knockout HeLa cell lysate (AB257181)

Homozygous : 1 bp deletion in exon1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1.

Disease

Adenocarcinoma

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Complementary Products

Assay Kits

AB139464

Cellular Calcineurin Phosphatase Activity Assay Kit (Colorimetric)

Functional Studies - Cellular Calcineurin Phosphatase Activity Assay Kit (Colorimetric) (AB139464)

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Primary Antibodies

AB282104

Anti-Calcineurin A antibody [EPR24997-22]

BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Calcineurin A antibody [EPR24997-22] (AB282104)

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Proteins & Peptides

AB128554

Recombinant Human Calcineurin A protein

SDS-PAGE - Recombinant Human Calcineurin A protein (AB128554)

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  • 0
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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
PPP3CA
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

Calcineurin A also known as PPP3CA is a highly specific protein phosphatase with a pivotal role in cellular signaling. It has a molecular mass of approximately 59 to 61 kDa depending on the isoform. Calcineurin A expression occurs in various tissues including the brain immune cells and cardiac tissues. It acts as the catalytic subunit of the enzyme calcineurin and is important for its function.
Biological function summary

Calcineurin A participates in numerous cellular processes through its phosphatase activity. As part of the calcineurin complex which includes a regulatory subunit it modulates the dephosphorylation of target proteins. This activity enables Calcineurin A to influence activation of T-lymphocytes by dephosphorylating the nuclear factor of activated T-cells (NFAT) a transcription factor responsible for immune response. Calcineurin A also plays a role in neuronal signaling and muscle function.

Pathways

Calcineurin A engages in the calcium signaling pathway which is vital for diverse cellular functions. It is related to proteins like calmodulin which binds calcium ions to activate Calcineurin A. Another important pathway includes the MAPK signaling pathway where Calcineurin A influences various cellular outcomes by regulating different transcription factors. Through these pathways Calcineurin A integrates signals that affect cellular growth survival and differentiation.

Calcineurin A is associated with cardiac hypertrophy and immunosuppressive conditions. In cardiac hypertrophy aberrant Calcineurin A activity leads to changes in heart muscle size and function. The protein also connects to the disorder of immunosuppression through its inhibition by drugs like cyclosporine which are used to prevent transplant rejection by targeting Calcineurin A. This inhibition blocks T-cell activation and provides therapeutic benefits in managing transplant patients.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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