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AB265717

Human ATP5E knockout HeLa cell line

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ATP5F1E KO cell line available to order. KO validated by. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.

View Alternative Names

ATP synthase H+ transporting mitochondrial F1 complex epsilon subunit, ATP synthase epsilon chain mitochondrial, ATP synthase subunit epsilon, ATP5E_HUMAN, ATPE, ATPase subunit epsilon, F(0)F(1) ATPase, H(+) transporting two sector ATPase, MGC104243, Mitochondrial ATP synthase epsilon chain, Mitochondrial ATPase, mitochondrial

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Sanger Sequencing - Human ATP5E knockout HeLa cell line (AB265717)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATP5E knockout HeLa cell line (AB265717)

Homozygous : 1 bp insertion in exon 1.

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1

Disease

Adenocarcinoma

Product details

Recommended control: Human wild-type HeLa cell line (ab255928). 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
ATP5F1E
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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.

ATP5E also known as ATP synthase epsilon subunit is a component of the ATP synthase enzyme complex specifically the F1 portion. This protein has an approximate molecular mass of 5.6 kDa. ATP5E is mainly present in the inner mitochondrial membrane where it plays a role in ATP production. The synthesis of ATP is a process important for cellular energy and ATP5E contributes by stabilizing the structure of the ATP synthase complex.
Biological function summary

ATP5E functions as an integral part of the ATP synthase complex which comprises multiple subunits. This complex is essential for oxidative phosphorylation a primary method through which cells produce ATP. ATP5E's role is structural helping to keep the F1 catalytic core properly aligned during the conversion of ADP and inorganic phosphate into ATP. Through this biological activity ATP5E supports energy metabolism necessary for the survival of cells.

Pathways

ATP5E participates in the oxidative phosphorylation pathway a critical process for cellular respiration. This pathway involves electron transport and proton pumping processes that generate the proton gradient ATP synthase exploits to synthesize ATP. Other proteins like ATP5C1 the gamma subunit are closely related to ATP5E in this pathway each contributing to the efficient production of ATP from ADP and inorganic phosphate.

ATP5E has connections with mitochondrial disorders such as Leigh syndrome. This disease results from mutations affecting mitochondrial function leading to severe energy deficits in cells. Mutations in ATP5E can disrupt the ATP synthase complex's function contributing to the pathology of these disorders. ATP5A1 another subunit of the ATP synthase complex also links with these diseases underlining the importance of all these components in maintaining mitochondrial integrity and function.

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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