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AB265980

Human ATG4D knockout HeLa cell line

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ATG4D 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

9830134P12Rik, APG4 D, APG4 autophagy 4 homolog D, APG4, S. cerevisiae, homolog of, D, ATG4 autophagy related 4 homolog D (S. cerevisiae), ATG4D_HUMAN, AUT-like 4 cysteine endopeptidase, AUTL4, Atg4dl, Autophagin-4, Autophagy-related cysteine endopeptidase 4, Autophagy-related protein 4 homolog D, Cysteine protease ATG4D, Cysteine protease involved in autophagy, MGC31226, autophagy 4, S. cerevisiae, homolog of, D, autophagy related 4D, cysteine peptidase

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Sanger Sequencing - Human ATG4D knockout HeLa cell line (AB265980)
  • Sanger seq

Unknown

Sanger Sequencing - Human ATG4D knockout HeLa cell line (AB265980)

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
ATG4D
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.

ATG4D also known as Autophagin-4 is a protein that plays a mechanical role in the autophagy process. It is an integral cysteine protease with a molecular mass of around 50 kDa. This protein is expressed across various tissues prominently in skeletal and cardiac muscle as well as in the nervous system. Its mechanical function involves the cleavage of LC3 proteins and other ATG8 family members which are essential steps for autophagosome maturation and function.
Biological function summary

The action of ATG4D facilitates the cellular autophagy process. It is not part of a larger protein complex but functions independently to regulate the recycling of cytoplasmic components. By ensuring the proper processing of LC3 ATG4D supports the degradation of damaged organelles and proteins maintaining cellular homeostasis. The protein's involvement in pathways promotes cellular survival during stress conditions such as nutrient deprivation.

Pathways

ATG4D primarily participates in the autophagy and mitophagy pathways. It is closely related to proteins such as ATG3 and ATG7 which coordinate to ensure the efficient conjugation and lipidation of ATG8 family members. In the mitophagy pathway it contributes to the selective degradation of mitochondria stabilizing mitochondrial quality control which is important for energy balance in cells.

ATG4D has connections to neurodegenerative diseases and cancer. Abnormal autophagy processes where ATG4D functions can contribute to conditions such as Parkinson's disease where defective mitophagy leads to neuronal cell death. In cancer the regulation of autophagy by ATG4D can influence tumor development and progression. Proteins such as p62/SQSTM1 linked to autophagy and parkin linked to mitophagy further demonstrate the role of ATG4D in these diseases indicating its importance in pathological conditions.

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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