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PRKN KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 77 bp deletion in exon 2.

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Images

Western blot - Human PARK2 (Parkin) knockout SH-SY5Y cell line (AB280042), expandable thumbnail
  • Sanger Sequencing - Human PARK2 (Parkin) knockout SH-SY5Y cell line (AB280042), expandable thumbnail

Key facts

Cell type
SH-SY5Y
Species or organism
Human
Tissue
Bone marrow
Form
Liquid
Knockout validation
Sanger Sequencing, Western blot
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 77 bp deletion in exon 2

Alternative names

AR JP, E3 ubiquitin ligase, E3 ubiquitin-protein ligase parkin, FRA6E, LPRS 2, PARK 2, PDJ, PRKN 2, PRKN2_HUMAN, Parkin 2, Parkinson disease (autosomal recessive juvenile) 2, Parkinson disease (autosomal recessive, juvenile) 2, parkin, Parkinson disease protein 2, Parkinson juvenile disease protein 2, Parkinson protein 2 E3 ubiquitin protein ligase, Parkinson protein 2, E3 ubiquitin protein ligase (parkin), Ubiquitin E3 ligase PRKN

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PRKN KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 77 bp deletion in exon 2.

Key facts

Cell type
SH-SY5Y
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, Homozygous: 77 bp deletion in exon 2
Disease
Neuroblastoma
Concentration
Loading...

Properties

Gene name
PRKN
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Zygosity
Homozygous

Quality control

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

Cell culture

Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female

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.
  • These cells grow as a mixture of floating and adherent cells.
  • Remove media containing floating cells and recover cells by centrifugation, detach cells using standard methods, combine with floating cells and transfer to a new culture flask.
  • A guide seeding density of 2x104 cells/cm2 is recommended.
  • Cells should be seeded at a density conducive to cell–cell communication to proliferate. If cells are seeded too sparsely, growth rate is reduced and cell death is high.
Culture medium
1:1 mixture of EMEM and F-12K + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

Storage

Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

Notes

Recommended control: Human wild-type SHSY-5Y cell line (ab275475). 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

The Parkin protein also known as PRK8 or Park2 is an E3 ubiquitin ligase with a molecular weight of approximately 52 kDa. This protein plays a critical role in tagging damaged proteins for degradation maintaining cellular health. Parkin is expressed in various tissues with significant levels in dopaminergic neurons in the brain. It is encoded by the PARK2 gene and has been linked to the regulation of mitochondrial quality and autophagy processes contributing to cellular homeostasis.

Biological function summary

Parkin is essential for the regulation of mitochondria through its involvement in the mitochondrial quality control system. It functions as part of a complex with other proteins that respond to mitochondrial damage by tagging them with ubiquitin molecules. This mechanism allows for the removal of defective mitochondria via mitophagy critical for preventing the accumulation of damaged cellular components.

Pathways

Parkin interacts with pathways involved in the cellular stress response particularly the PINK1 (PTEN Induced Kinase 1) pathway. PINK1 phosphorylates Parkin activating it to label damaged mitochondria. Another critical pathway involves proteasomal degradation where Parkin collaborates with Ubiquitin to manage protein turnover. These pathways highlight its relationships with other cellular stress-regulating proteins enhancing our understanding of its roles in maintaining cellular integrity.

Associated diseases and disorders

Mutations in the gene coding for Parkin are linked to Parkinson's disease (PD) and some forms of juvenile autosomal recessive parkinsonism. The Parkin protein's dysfunctional activity leads to impaired mitochondrial management and protein aggregation in neurons contributing significantly to neurodegenerative disease. In conditions such as PD Parkin interacts with other proteins such as PINK1 reinforcing its role in mitochondrial protection and indicating the protein's importance in disease progression and potential therapeutic targeting.

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