Human PPM1A knockout HeLa cell line
- Advanced Validation
- What is this?
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PPM1A KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 4 bp deletion in exon 3 and Insertion of the selection cassette in exon 3. 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
EC 3.1.3.16, FLJ42306, IA, MGC9201, Mpp alpha, PP2C-alpha, PP2CA, PPM1A_HUMAN, PPPM1A, Protein phosphatase 1A, Protein phosphatase 1A (formerly 2C) magnesium dependent alpha isoform, Protein phosphatase 1A magnesium dependent alpha, Protein phosphatase 2C alpha, Protein phosphatase 2C alpha isoform, Protein phosphatase 2C isoform alpha, Protein phosphatase IA, Protein phosphatase, Mg2+/Mn2+ dependent, 1A
- WB
Lab
Western blot - Human PPM1A knockout HeLa cell line (AB265348)
Lanes 1-4 : Merged signal (red and green). Green - ab14824 observed at 42 kDa. Red - loading control ab181602 observed at 36 kDa.
ab14824 Anti-PPM1A antibody [p6c7] was shown to specifically react with PPM1A in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265348 (knockout cell lysate ab259055) was used. Wild-type and PPM1A knockout samples were subjected to SDS-PAGE. ab14824 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-PPM1A antibody [p6c7] (<a href='/en-us/products/primary-antibodies/ppm1a-antibody-p6c7-ab14824'>ab14824</a>) at 1/500 dilution
Lane 1:
Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
PPM1A knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 2:
Western blot - Human PPM1A knockout HeLa cell line (ab265348)
Lane 3:
HAP1 whole cell lyate at 20 µg
Lane 4:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-680rd-preadsorbed-ab216777'>ab216777</a>) at 1/10000 dilution
Predicted band size: 41 kDa,42 kDa
Observed band size: 42 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human PPM1A knockout HeLa cell line (AB265348)
Allele-1 : 4 bp deletion in exon 3.
- Sanger seq
Unknown
Sanger Sequencing - Human PPM1A knockout HeLa cell line (AB265348)
Allele-2 : Insertion of the selection cassette in exon 3.
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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
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
PPM1A regulates many cellular mechanisms by dephosphorylating targets and acting as a negative regulator of several signaling pathways. It modulates stress responses cell cycle progression and immune responses. PPM1A does not typically form complexes with other proteins but interacts closely with specific signaling molecules to control their activity. This regulation influences cellular homeostasis and growth in response to external and internal signals.
Pathways
PPM1A influences the TGF-beta signaling pathway where it interacts with and deactivates SMAD proteins by dephosphorylation. It also participates in the p38 MAPK pathway. These interactions highlight its role in controlling transduction signals initiated by growth factors and stress. By regulating these pathways PPM1A maintains a balance in cellular responses to growth and environmental stresses.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPM1A antibody [p6c7] (AB14824)](https://content.abcam.com/products/images/ppm1a-antibody-p6c7-ab14824--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img73582.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com