Human PARK2 (Parkin) knockout SH-SY5Y cell line
- Advanced Validation
- What is this?
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PRKN KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 77 bp deletion in exon 2. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View 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
- WB
Lab
Western blot - Human PARK2 (Parkin) knockout SH-SY5Y cell line (AB280042)
Lanes 1 - 4 : Merged signal (red and green). Green - ab77924 observed at 49 kDa. Red - loading control ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37 kDa.
ab77924 was shown to react with Parkin in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PRKN knockout cell line ab280042 (PRKN knockout cell lysate ab280101). Wild-type SH-SY5Y and PRKN knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab77924 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4 °C at 5 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes:
Western blot - Anti-Parkin antibody [PRK8] (<a href='/en-us/products/primary-antibodies/parkin-antibody-prk8-ab77924'>ab77924</a>) at 5 µg/mL
Lane 1:
Wild-type SH-SY5Y cell lysate at 20 µg
Lane 2:
PRKN knockout SH-SY5Y cell lysate at 20 µg
Lane 2:
Western blot - Human PARK2 (Parkin) knockout SH-SY5Y cell line (ab280042)
Lane 3:
Human Brain tissue lysate at 20 µg
Lane 4:
HUVEC cell lysate at 20 µg
Predicted band size: 52 kDa
Observed band size: 49 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human PARK2 (Parkin) knockout SH-SY5Y cell line (AB280042)
Human PRKN KO in SH-SY5Y Cells with 77 bp Deletion in Exon 2
Reactivity data
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?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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
- 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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Western blot - Anti-Parkin antibody [PRK8] (AB77924)](https://content.abcam.com/products/images/parkin-antibody-prk8-ab77924--western-blot-img168393.jpg)
Primary Antibodies
AB179483
Anti-HIF-1 alpha antibody [EPR16897]
RabMAb|Advanced Validation|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [EPR16897] (AB179483)](https://content.abcam.com/products/images/hif-1-alpha-antibody-epr16897-ab179483--immunocytochemistry-immunofluorescence-img85620.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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