Skip to main content

PINK1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9,
7 bp Deletion in Exon 1.

Be the first to review this product! Submit a review

Images

Sanger Sequencing - Human PINK1 knockout SH-SY5Y cell line (AB280876), expandable thumbnail
  • Western blot - Human PINK1 knockout SH-SY5Y cell line (AB280876), expandable thumbnail

Publications

Key facts

Cell type
SH-SY5Y
Species or organism
Human
Tissue
Bone marrow
Form
Liquid
Knockout validation
Sanger Sequencing
Mutation description
Knockout achieved by using CRISPR/Cas9, 7 bp Deletion in Exon 1.

Alternative names

BRPK, FLJ27236, PARK 6, PINK1_HUMAN, PTEN induced putative kinase 1, PTEN-induced putative kinase protein 1, Phosphatase and Tensin Homolog, Protein kinase BRPK, Serine/threonine kinase PINK1 mitochondrial, Serine/threonine protein kinase PINK1 mitochondrial, Serine/threonine-protein kinase PINK1, mitochondrial

Recommended products

PINK1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9,
7 bp Deletion in Exon 1.

Key facts

Cell type
SH-SY5Y
Form
Liquid
Mutation description
Knockout achieved by using CRISPR/Cas9, 7 bp Deletion in Exon 1.
Disease
Neuroblastoma
Concentration
Loading...

Properties

Gene name
PINK1
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing

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 PTEN-induced kinase 1 (PINK1) is a serine/threonine-protein kinase with a molecular weight of approximately 63 kDa. It plays a significant role in mitochondrial quality control through its kinase activity. PINK1 gets expressed in various tissues with a high presence in the brain. The protein localizes to the outer membrane of damaged mitochondria to initiate mitophagy an important cellular process for clearing dysfunctional mitochondria.

Biological function summary

The PINK1 protein detects mitochondrial damage and recruits Parkin an E3 ubiquitin ligase to the damaged mitochondria. This interaction leads to the ubiquitylation of mitochondrial substrates and triggers their degradation. PINK1 Parkin and other proteins form a complex that facilitates the removal of damaged mitochondria through autophagy. This mechanism ensures cellular health and energy balance by maintaining a pool of functional mitochondria.

Pathways

PINK1 integrates into the mitochondrial quality control and mitophagy pathways. It has a fundamental role in the PINK1-Parkin pathway which is critical for maintaining mitochondrial integrity. In this pathway PINK1 phosphorylates both ubiquitin and Parkin enhancing Parkin’s E3 ligase activity. Another pathway that PINK1 participates in is the PTEN-induced kinase pathway where it regulates mitochondrial dynamics and homeostasis via cross-talk with other mitochondrial proteins such as DJ-1 and LRRK2.

Associated diseases and disorders

PINK1 mutations are strongly associated with familial Parkinson’s disease attributing to its role in preserving neuronal function through mitochondrial regulation. Deficient PINK1 function leads to accumulation of damaged mitochondria contributing to neuronal cell death. Additionally PINK1 dysfunction is implicated in Alzheimer’s disease as impaired mitochondrial clearance is a contributing factor. Here the PINK1 interaction with other proteins like Parkin and DJ-1 highlights its importance in neurodegenerative disorders.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

2 product images

  • Sanger Sequencing - Human PINK1 knockout SH-SY5Y cell line (ab280876), expandable thumbnail

    Sanger Sequencing - Human PINK1 knockout SH-SY5Y cell line (ab280876)

    Human PINK1 KO in SH-SY5Y Cells with 7 bp Deletion in Exon 1.

  • Western blot - Human PINK1 knockout SH-SY5Y cell line (ab280876), expandable thumbnail

    Western blot - Human PINK1 knockout SH-SY5Y cell line (ab280876)

    Western blot: Anti-PINK1 antibody [MJF-R32-7] (Anti-PINK1 antibody [MJF-R32-7] ab300623) staining at 1/1000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta.

    In Western blot, Anti-PINK1 antibody [MJF-R32-7] ab300623 was shown to bind specifically to PINK1. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times before development with a high-sensitivity ECL substrate kit and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

    In Western blot, Anti-PINK1 antibody [MJF-R32-7] ab300623 was shown to bind specifically to PINK1. A band was observed at 63kDa in wild-type SH-SY5Y treated with CCCP cell lysates with no signal observed at this size in PINK1 knockout cell line (ab280876).

    All lanes: Western blot - Anti-PINK1 antibody [MJF-R32-7] (Anti-PINK1 antibody [MJF-R32-7] ab300623)

    Lane 1: Wild-type SH-SY5Y treated CCCP, CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229 (10 μM, 24 h) cell lysate at 20 µg

    Lane 2: Wild-type SH-SY5Y control CCCP (0 μM, 24 h) cell lysate at 20 µg

    Lane 3: PINK1 knockout SH-SY5Y (ab280876) treated CCCP, CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229 (10 uM, 24 h) cell lysate at 20 µg

    Lane 4: Western blot - Human PINK1 knockout SH-SY5Y cell line (ab280876) at 20 µg

    Lane 5: Wild-type HEK-293 Treated CCCP, CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229 (10 μM, 24 h) cell lysate at 20 µg

    Lane 6: PINK1 knockout HEK-293 Treated CCCP, CCCP, Mitochondrial oxidative phosphorylation uncoupler ab141229 (10 μM, 24 h) cell lysate at 20 µg

    Secondary

    All lanes: HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 63 kDa

    Observed band size: 63 kDa

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com