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AB261816

Human CAT (Catalase) knockout HeLa cell line

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CAT 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 1 bp insertion in exon 2.

View Alternative Names

CAS1, CATA_HUMAN, CS 1, Catalase, MGC138422, MGC138424

2 Images
Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell line (AB261816)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell line (AB261816)

Allele-1 : 1 bp deletion in exon 2.

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell line (AB261816)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell line (AB261816)

Allele-2 : 1 bp insertion in exon 2.

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

Antibiotic resistance

Puromycin 1µg/mL

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
CAT
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.

Catalase also known as CAT is an enzyme that catalyzes the decomposition of hydrogen peroxide into water and oxygen. This enzyme has a molecular weight of approximately 240 kDa and typically forms a tetramer. Catalase mainly resides in peroxisomes functioning as a peroxisome marker. It expresses abundantly in the liver kidneys and erythrocytes where it plays significant roles in cellular protection against oxidative damage. The presence of Catalase makes it an ideal candidate for use in peroxisome staining and peroxisome image analysis in research.
Biological function summary

Catalase contributes to antioxidant defense by breaking down hydrogen peroxide preventing cellular damage. In peroxisomes it works alongside other peroxisomal enzymes to maintain cell health and metabolic regulation. Catalase does not form a complex but interacts closely with other enzymes like superoxide dismutase which dismutates superoxide radicals into less harmful substances. Increased Catalase activity levels can be measured using Catalase activity kits and activity assays allowing us to learn about peroxisome function.

Pathways

Hydrogen peroxide removal by Catalase is vital in the reactive oxygen species (ROS) metabolic process and plays a part in the cellular response to oxidative stress. Catalase interacts with the glutathione peroxidase pathway safeguarding cells from oxidative stress-related damage. Superoxide dismutase works synergistically with Catalase transforming superoxide anions into hydrogen peroxide before its decomposition by Catalase. These activities highlight the essential role Catalase plays in protecting cells from oxidative stress damage.

Catalase relates to conditions like acatalasemia and diabetes. Acatalasemia a condition caused by a deficiency of Catalase increases the risk of developing diabetes and other oxidative stress-related diseases. Mutations in the CAT gene can lead to decreased Catalase activity contributing to the onset of these conditions. Additionally Catalase works with other proteins like glutathione peroxidase in mitigating the effects of oxidative stress with deficiencies potentially exacerbating complications in diabetes management.

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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