Human PPP3CA (Calcineurin A) knockout HeLa cell line
- Advanced Validation
- What is this?
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(1 Publication)
PPP3CA KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 1. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
Alpha isoform formerly PPP2B, CALN, CALNA, CALNA 1, CAM-PRP catalytic subunit, CNA, CNA 1, CNA alpha, Calcineurin A alpha, Calcineurin A1, CalcineurinA, Calmodulin-dependent calcineurin A subunit alpha isoform, PP2BA_HUMAN, PPP2B, Ppp3ca, Protein phosphatase 2B catalytic subunit, Protein phosphatase 3 (formerly 2B) catalytic subunit alpha isoform, Protein phosphatase 3 catalytic subunit alpha isoform PPP3CA, Protein phosphatase 3 catalytic subunit alpha isozyme, Serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform
- WB
Lab
Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell line (AB265130)
Lanes 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° 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.
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 (ab265130)
Predicted band size: 59 kDa
Observed band size: 59 kDa
false
- WB
Lab
Western blot - Human PPP3CA (Calcineurin A) knockout HeLa cell line (AB265130)
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° 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.
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 (ab265130)
Predicted band size: 59 kDa
Observed band size: 59 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PPP3CA (Calcineurin A) knockout HeLa cell line (AB265130)
Homozygous : 1 bp deletion in exon 1.
- Cell Culture
Unknown
Cell Culture - Human PPP3CA (Calcineurin A) knockout HeLa cell line (AB265130)
Representative images of PPP3CA knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
BMC cancer 24:508 PubMed39103807
2024
Applications
Unspecified application
Species
Unspecified reactive species
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