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AB256859

Human CAT (Catalase) knockout HeLa cell lysate

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CAT KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 4 bp deletion in exon1 and Insertion of the selection cassette in exon1.

View Alternative Names

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

5 Images
Western blot - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)
  • WB

Lab

Western blot - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : CAT knockout HeLa cell lysate (20 μg)

Lanes 1-2 : Merged signal (red and green). Green - ab76024 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

ab76024 Anti-Catalase antibody [EP1929Y] - Peroxisome Marker was shown to specifically react with Catalase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265250 (knockout cell lysate ab256859) was used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab76024 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Catalase antibody [EP1929Y] - Peroxisome Marker (<a href='/en-us/products/primary-antibodies/catalase-antibody-ep1929y-peroxisome-marker-ab76024'>ab76024</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CAT knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CAT (Catalase) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cat-catalase-knockout-hela-cell-line-ab265250'>ab265250</a>)

Predicted band size: 60 kDa

Observed band size: 60 kDa

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Western blot - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)
  • WB

Lab

Western blot - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)

Lane 1 : Wild-type HeLa cell lysate (20 μg)

Lane 2 : CAT knockout HeLa cell lysate (20 μg)

Lanes 1-2 : Merged signal (red and green). Green - ab209211 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

ab209211 Anti-Catalase antibody [EPR20198] was shown to specifically react with Catalase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265250 (knockout cell lysate ab256859) was used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab209211 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Catalase antibody [EPR20198] - Peroxisome Marker (<a href='/en-us/products/primary-antibodies/catalase-antibody-epr20198-peroxisome-marker-ab209211'>ab209211</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CAT knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CAT (Catalase) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cat-catalase-knockout-hela-cell-line-ab265250'>ab265250</a>)

Predicted band size: 60 kDa

Observed band size: 60 kDa

false

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)

Allele-1 : 4 bp deletion in exon1

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)

Allele-3 : Insertion of the selection cassette in exon1

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)
  • Sanger seq

Unknown

Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell lysate (AB256859)

Allele-2 : 1 bp deletion in exon1

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 4 bp deletion in exon1 and Insertion of the selection cassette in exon1.

Disease

Adenocarcinoma

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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.

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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, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Primary Antibodies

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Primary Antibodies

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Anti-Catalase antibody [EPR20198] - Peroxisome Marker

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Product promise

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