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AB301175

Human PPM1D knockout A549 cell line

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PPM1D KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control available. 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, PP2C-delta, PP2CD, PPM1D_HUMAN, Protein phosphatase 1D, Protein phosphatase 2C delta isoform, Protein phosphatase 2C isoform delta, Protein phosphatase Mg2+/Mn2+ dependent 1D, Protein phosphatase Wip1, Protein phosphatase magnesium-dependent 1 delta, WIP 1, Wild type p53 induced phosphatase 1, p53-induced protein phosphatase 1, protein phosphatase 1D magnesium-dependent delta isoform

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Next Generation Sequencing - Human PPM1D knockout A549 cell line (AB301175)
  • NGS

Lab

Next Generation Sequencing - Human PPM1D knockout A549 cell line (AB301175)

184 bp deletion after Ser 168 (allele 1); 130 bp deletion after Leu 166 (allele 2); 172 bp deletion and 121 bp insertion after Trp 171 (allele 3) of WT protein

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Disease

Carcinoma

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.

Recommended control: Human wild-type A549 cell line (ab288558). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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.

Properties and storage information

Gene name
PPM1D
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.
  • Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium

F-12K + 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.

The PPM1D protein also known as wild-type p53-induced phosphatase 1 (WIP1) is a serine/threonine phosphatase. Its molecular weight is approximately 66 kDa. PPM1D is an enzyme expressed in various tissues including the brain heart and kidney. This phosphatase plays an important role in the regulation of the cellular stress response by dephosphorylating specific substrates.
Biological function summary

PPM1D/WIP1 acts as a modulator of cellular stress and survival mechanisms. The protein influences the DNA damage response and supports cell cycle progression by deactivating critical checkpoint proteins such as p38 MAPK and p53. It does not form part of a larger protein complex but interacts directly with these proteins to exert its biological functions.

Pathways

PPM1D engages with mechanisms linked to the DNA damage response and the p53 signaling pathway. It counteracts the activation of p53 a protein integral in cell cycle arrest and apoptosis which helps cells to recover post-DNA damage. PPM1D also interacts with proteins like ATM and CHK2 which are part of the DNA repair pathway playing a role in cellular recovery and survival.

PPM1D links to cancer development and progression. Its role in deactivating p53 and facilitating continued cell proliferation makes it relevant to tumorigenesis. Also PPM1D's connection to ATM and the DNA damage response becomes significant in neurodegenerative diseases. Dysregulation of PPM1D can contribute to an improper cellular response to genomic instability contributing to disease pathogenesis.

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

Product protocols

Product promise

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