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AB265691

Human SMEK2 knockout HeLa cell line

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PPP4R3B 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 1. 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 SMEK2 knockout HeLa cell line (AB265691)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMEK2 knockout HeLa cell line (AB265691)

Homozygous : 2 bp deletion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 1

Disease

Adenocarcinoma

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
PPP4R3B
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.

SMEK2 also known as protein phosphatase 4 regulatory subunit 3B (PPP4R3B) is a regulatory subunit of the protein phosphatase 4 (PP4) complex. SMEK2 exhibits a molecular mass of about 106 kDa. It is distributed in various tissues with notable expression in the brain and skeletal muscle. As part of the PP4 complex its primary role is to guide and regulate the specific functions of the catalytic subunits of the complex.
Biological function summary

SMEK2 regulates cellular processes by modulating phosphatase activity. It acts as part of the PP4 complex which includes the PP4 catalytic subunit and other regulatory subunits. This complex plays a role in diverse biological functions such as cellular response to DNA damage and cell division. By affecting dephosphorylation events SMEK2 influences cellular signaling allowing for precise cellular responses under varying conditions.

Pathways

SMEK2 significantly impacts the DNA damage response and cell cycle regulation pathways. It interacts closely with proteins such as PP4C the catalytic subunit of the PP4 complex and has connections to key checkpoint regulators in these pathways. Such interactions ensure accurate DNA repair and proper progression through cell cycle phases. Changes in SMEK2's functionality or expression can disrupt these pathways leading to improper cellular responses.

Altered SMEK2 activity or expression has been associated with cancer and neurodegenerative diseases. In cancer SMEK2 along with PP4C influences tumor suppressor pathways by regulating processes like DNA damage repair. Disruption in these interactions can lead to uncontrolled cell proliferation. Additionally abnormalities in SMEK2 have been linked to Alzheimer's disease possibly due to its involvement in neuronal signaling pathways and response to cellular stress.

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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