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AB266118

Human MAPKAPK5 (PRAK/MK5) knockout HEK-293T cell line

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MAPKAPK5 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 2 bp deletion in exon 5 and 4 bp deletion in exon 5. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 Images
Sanger Sequencing - Human MAPKAPK5 (PRAK/MK5) knockout HEK-293T cell line (AB266118)
  • Sanger seq

Unknown

Sanger Sequencing - Human MAPKAPK5 (PRAK/MK5) knockout HEK-293T cell line (AB266118)

Allele-1 : 4 bp deletion in exon5

Sanger Sequencing - Human MAPKAPK5 (PRAK/MK5) knockout HEK-293T cell line (AB266118)
  • Sanger seq

Unknown

Sanger Sequencing - Human MAPKAPK5 (PRAK/MK5) knockout HEK-293T cell line (AB266118)

Allele-2 : 2 bp deletion in exon 5.

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, 2 bp deletion in exon 5 and 4 bp deletion in exon 5

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
MAPKAPK5
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger 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.
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.

PRAK also known as MK5 is a serine/threonine protein kinase with a molecular weight of approximately 54 kDa. It is expressed in various tissues like the heart brain liver and skeletal muscle. PRAK belongs to the mitogen-activated protein kinase (MAPK) signaling pathway and gets activated by phosphorylation. Mechanically PRAK interacts with and phosphorylates other proteins which affects their activity and function playing an important role in regulating cellular processes such as stress response and the cell cycle.
Biological function summary

PRAK/MK5 is a critical component in the regulation of cellular responses to stress stimuli. It is a participant in the activation loop of the MAPK p38 signaling pathway where it serves to influence the phosphorylation state and activity of downstream effectors. PRAK is not typically known to be part of a large complex but it interacts directly with other kinases to amplify or dampen the stress response signal. This allows cells to adapt to varied stress conditions by modulating gene expression apoptosis and other cell fate decisions.

Pathways

PRAK/MK5 plays essential roles within the p38 MAPK and ERK signaling pathways. It closely associates with p38 MAPK which phosphorylates and activates PRAK enabling it to further disseminate the signal. In these pathways PRAK interplays with proteins such as MAPKAPK-2 and HSP27 facilitating diverse cellular activities like inflammation and differentiation. These signaling networks allow PRAK to serve as a link between external stimuli and internal cellular outcomes.

PRAK/MK5 influences cancer and cardiovascular diseases. Its activity connects closely with tumor suppression wherein it regulates actions involving proteins like p53 and HSP27 impacting cell cycle arrest and apoptosis. In cardiovascular contexts altered PRAK function can relate to heart disease since it regulates stress-induced cardiac responses. Understanding PRAK's interplay with these proteins sheds light on its potential as a therapeutic target for treating diseases related to stress signaling dysfunction.

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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