Human CAT (Catalase) knockout HeLa cell line
- Advanced Validation
- What is this?
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CAT KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 1 and 4 bp deletion in exon 1 and Insertion of the selection cassette in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
CAS1, CATA_HUMAN, CS 1, Catalase, MGC138422, MGC138424
- WB
Lab
Western blot - Human CAT (Catalase) knockout HeLa cell line (AB265250)
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] - 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. ab209211 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 (ab265250)
Predicted band size: 60 kDa
Observed band size: 60 kDa
false
- WB
Lab
Western blot - Human CAT (Catalase) knockout HeLa cell line (AB265250)
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° 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 (ab265250)
Predicted band size: 60 kDa
Observed band size: 60 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell line (AB265250)
Allele-1 : 4 bp deletion in exon 1.
- Sanger seq
Unknown
Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell line (AB265250)
Allele-3 : Insertion of the selection cassette in exon 1.
- Cell Culture
Unknown
Cell Culture - Human CAT (Catalase) knockout HeLa cell line (AB265250)
Representative images of CAT knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa 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 seq
Unknown
Sanger Sequencing - Human CAT (Catalase) knockout HeLa cell line (AB265250)
Allele-2 : 1 bp deletion in exon 1.
Reactivity data
Product details
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?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
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.
Quality control
STR analysis
TH01, D16S539, TPOX, CSF1PO, D13S317, D7S820, D5S818
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Primary Antibodies
AB209211
Anti-Catalase antibody [EPR20198] - Peroxisome Marker
KO Validated|RabMAb|Recombinant|Lab Essentials|20ul selling size
![Flow Cytometry (Intracellular) - Anti-Catalase antibody [EPR20198] - Peroxisome Marker (AB209211)](https://content.abcam.com/products/images/catalase-antibody-epr20198-peroxisome-marker-ab209211--flow-cytometry-intracellular-img80369.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com