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AB274925

Human ATP13A2 knockout HEK-293 cell line

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ATP13A2 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 99%.

View Alternative Names

AT132_HUMAN, ATPase type 13A2, CLN12, FLJ26510, HSA9947, KRPPD, PARK9, Probable cation-transporting ATPase 13A2, Putative ATPase, RP1-37C10.4

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Next Generation Sequencing - Human ATP13A2 knockout HEK-293 cell line (AB274925)
  • NGS

Supplier Data

Next Generation Sequencing - Human ATP13A2 knockout HEK-293 cell line (AB274925)

1 bp deletion after Ser216 of the WT protein

Next Generation Sequencing - Human ATP13A2 knockout HEK-293 cell line (AB274925)
  • NGS

Supplier Data

Next Generation Sequencing - Human ATP13A2 knockout HEK-293 cell line (AB274925)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 99%

Key facts

Cell type

HEK-293

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Next Generation Sequencing

Mutation description

Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 99%

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

Product details

Recommended control: Human wild-type HEK-293 cell line (ab259776). 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.

What's included?

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Properties and storage information

Gene name
ATP13A2
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation 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.

ATP13A2 also known as PARK9 is a lysosomal P5-type ATPase with a molecular mass of approximately 163 kDa. It functions primarily as a cation transporter involved in the transport of divalent metal ions such as manganese and zinc across lysosomal membranes. The ATP13A2 protein is expressed in various tissues with high levels noted in the brain particularly within the neurons of the substantia nigra. The presence of ATP13A2 is also confirmed in kidney tissues and immune cells indicating a wide tissue distribution.
Biological function summary

ATP13A2 plays a significant role in maintaining metal ion homeostasis inside cells. It is an essential component of the complex regulatory system that ensures proper lysosomal function. The protein impacts autophagy by influencing lysosomal biogenesis and integrity. Furthermore its function in metal ion transport and detoxification suggests a protective role for neurons against metal-induced oxidative stress.

Pathways

ATP13A2 is an important participant in the lysosomal degradation pathway and the autophagy-lysosome pathway. It interacts with proteins such as alpha-synuclein known for its role in neurodegenerative diseases and cellular stress responses. In the context of metal ion homeostasis ATP13A2 operates alongside other transporters and enzymes to maintain cellular ion balance and prevent toxicity.

ATP13A2 mutations associate strongly with neurodegenerative conditions like Parkinson's disease (PD) and Kufor-Rakeb syndrome a rare juvenile-onset parkinsonism. Disruption in ATP13A2 activity leads to cellular dysfunction and neurodegeneration due to impaired lysosomal degradation and metal ion imbalance. It is connected to PARK7 (DJ-1) another protein linked to the pathogenesis of PD suggesting shared pathways or compensatory mechanisms between these proteins in neuronal survival and pathology.

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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