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AB266310

Human PPM1B knockout HEK-293T cell line

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PPM1B KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Sanger Sequencing - Human PPM1B knockout HEK-293T cell line (AB266310)
  • Sanger seq

Unknown

Sanger Sequencing - Human PPM1B knockout HEK-293T cell line (AB266310)

Homozygous : 2 bp deletion in exon 3

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 3

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?

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Properties and storage information

Gene name
PPM1B
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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.
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.

PPM1B also known as protein phosphatase 1B is a serine/threonine phosphatase with a molecular mass of approximately 55 kDa. This protein plays an important role in dephosphorylating serine/threonine residues on its target proteins adjusting their functional states. PPM1B is widely expressed in various tissues throughout the body including the heart skeletal muscle and liver. Its activity helps in modulating different signaling pathways by reversing the phosphorylation status of target proteins.
Biological function summary

PPM1B influences diverse cellular processes such as cell cycle regulation and stress response. It is not part of a large complex but interacts with specific substrates to exert its effects. PPM1B can impact apoptosis and cell differentiation through its action on certain substrates. Its regulatory function is important for maintaining cellular homeostasis and adaptation to environmental changes.

Pathways

Multiple signaling cascades demonstrate the involvement of PPM1B. It plays a significant role in the regulation of the p38 MAPK pathway which is important for stress response and inflammatory processes. In this pathway PPM1B interacts with MAPKAPK2 modulating its activity. PPM1B also participates in the TGF-beta signaling pathway where it can affect the phosphorylation status of SMADs. These interactions emphasize PPM1B's regulatory importance in response to extracellular signals.

Alterations in PPM1B activity are linked to cardiovascular and inflammatory conditions. For example changes in its function can contribute to the pathogenesis of hypertension through its effect on the TGF-beta pathway and related proteins like SMAD2. In inflammatory diseases the role of PPM1B in the p38 MAPK pathway shows its connection to proteins such as MAPKAP2 highlighting its potential involvement in inflammatory responses. These connections demonstrate the therapeutic interest in targeting PPM1B for disease treatment and management.

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

Product protocols

Product promise

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