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AB267307

Human AK3 knockout HEK-293T cell line

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AK3 KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and 7 bp deletion in exon 2.

View Alternative Names

AK3, formerly, AK4, AK4, mouse, homolog of, AKL3L, AKL3L1, Adenylate kinase 3, Adenylate kinase 3 alpha-like 1, Adenylate kinase 3, formerly, FIX, GTP:AMP phosphotransferase, GTP:AMP phosphotransferase, mitochondrial, KAD3_HUMAN, OTTHUMP00000021015, OTTHUMP00000021016, RP11 6J24.4, adenylate kinase 6, adenylate kinase 3 like 1, mitochondrial

3 Images
Cell Culture - Human AK3 knockout HEK-293T cell line (AB267307)
  • Cell Culture

Unknown

Cell Culture - Human AK3 knockout HEK-293T cell line (AB267307)

Representative images of AK3 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.

Sanger Sequencing - Human AK3 knockout HEK-293T cell line (AB267307)
  • Sanger seq

Unknown

Sanger Sequencing - Human AK3 knockout HEK-293T cell line (AB267307)

Allele-1 : 7 bp deletion in exon2

Sanger Sequencing - Human AK3 knockout HEK-293T cell line (AB267307)
  • Sanger seq

Unknown

Sanger Sequencing - Human AK3 knockout HEK-293T cell line (AB267307)

Allele-2 : 1 bp deletion in exon 2.

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, 1 bp deletion in exon 2 and 7 bp deletion in exon 2

Product details

Recommended control: Human wild-type HEK293T cell line (ab255449). 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.

What's included?

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Properties and storage information

Gene name
AK3
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.

AK3 also known as adenylate kinase 3 is a mitochondrial enzyme involved in energy metabolism. It has a molecular mass of approximately 26 kDa and is mainly found in the mitochondrial matrix. AK3 catalyzes the reversible transfer of phosphate groups among adenosine phosphates which plays a role in cellular energy homeostasis. Expression of AK3 occurs in tissues with high metabolic rates such as the heart liver and muscle reflecting its importance in energy demand areas.
Biological function summary

Adenylate kinase 3 facilitates the maintenance of cellular energy balance by ensuring the proper concentration of adenine nucleotides within the mitochondria. This enzyme is an important component of the adenine nucleotide translocase complex which regulates nucleotide transport across the inner mitochondrial membrane. Through its activity AK3 contributes to the recycling of ATP and ADP supporting overall energy metabolism and mitochondrial function.

Pathways

Adenylate kinase 3 plays a significant role in the purine nucleotide cycle and oxidative phosphorylation pathways. In the purine nucleotide cycle it interacts with enzymes like nucleoside diphosphate kinase for maintaining proper levels of ADP and ATP. Within oxidative phosphorylation AK3 helps in balancing energy supply in concert with other mitochondrial proteins like ATP synthase ensuring efficient ATP production and utilization in energy-intensive processes.

Alterations in adenylate kinase 3 function have been linked to mitochondrial diseases and cardiac disorders. Mitochondrial dysfunction resulting from impaired AK3 activity can lead to energy deficiency contributing to diseases like Leigh syndrome. Similarly inadequate AK3 function may affect cardiac energy metabolism relating to heart failure conditions where proper energy supply is essential. AK3's interactions with proteins like creatine kinase further emphasize its role in maintaining cardiac energy stability.

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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