CTNNB1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Images
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
Alternative names
Beta-catenin, CATNB, CHBCAT, CTNB1_HUMAN, CTNNB, CTNNB1, Cadherin-associated protein, Catenin (cadherin associated protein), beta 1, 88kDa, Catenin beta-1, DKFZp686D02253, FLJ25606, FLJ37923, OTTHUMP00000162082, OTTHUMP00000165222, OTTHUMP00000165223, OTTHUMP00000209288, OTTHUMP00000209289, b-catenin
Recommended products
CTNNB1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
Key facts
- Cell type
- MCF7
- Species or organism
- Human
- Tissue
- Breast
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing, Western blot
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- CTNNB1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- MEM + 10% FBS + 0.01 mg/ml bovine insulin
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type MCF7 cell line (ab288560). 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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Beta Catenin also known by names such as CTNNB1 or beta-chip is an important protein involved in cell signaling and adhesion. This protein has a molecular weight of around 88 kDa. Beta Catenin is expressed in many cell types and tissues indicating its widespread role in various biological processes. It functions mechanically by mediating the linkage between cadherins and the actin cytoskeleton facilitating cell-cell adhesion. Beta Catenin is also a central part of transcription regulation processes in the nucleus.
- Biological function summary
This protein plays roles in both cell adhesion and the regulation of gene expression. Beta Catenin is a critical component of the Wnt signaling pathway where it can form complexes with other proteins to influence gene transcription. In the absence of Wnt signaling beta Catenin levels are low due to its degradation. However when the pathway is active it accumulates in the cytoplasm and eventually translocates to the nucleus where it interacts with TCF/LEF transcription factors to regulate the expression of target genes.
- Pathways
Beta Catenin plays a central role in the Wnt signaling pathway and influences cell fate decisions and cellular proliferation. It acts in concert with proteins such as Dishevelled (DVL) and Axin to coordinate these important biological processes. In the absence of Wnt signaling proteins such as APC and GSK-3β are responsible for beta Catenin degradation keeping its cellular levels in check. Beta Catenin’s interaction with transcription factors in the nucleus makes it pivotal in the regulation of cell and tissue homeostasis.
- Associated diseases and disorders
Beta Catenin has associations with colorectal cancer and hepatocellular carcinoma. Its dysregulation can lead to unchecked cell proliferation and tumorigenesis. Often mutations in the beta Catenin gene (CTNNB1) or components of the Wnt pathway like APC are implicated in the development of these cancers. Its interplay with E-cadherin is important for maintaining tissue architecture and disruptions can lead to invasive cancer phenotypes. Understanding beta Catenin’s role provides insights into therapeutic strategies for these cancers.
Product promise
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Next Generation Sequencing - Human CTNNB1 knockout MCF7 cell line (ab286762)
216 bp deletion after the 91st AA of the WT protein
Western blot - Human CTNNB1 knockout MCF7 cell line (ab286762)
False colour image of Western blot: Anti-beta Catenin antibody [E247] - ChIP Grade staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta Catenin antibody [E247] - ChIP Grade ab32572 was shown to bind specifically to beta Catenin. A band was observed at 85/90 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CTNNB1 knockout cell line ab286762. To generate this image, wild-type and CTNNB1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (Anti-beta Catenin antibody [E247] - ChIP Grade ab32572) at 1/5000 dilution
Lane 1: Wild-type MCF7 cell lysate at 20 µg
Lane 2: CTNNB1 knockout MCF7 cell lysate, at 20 µg
Lane 3: HeLa cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 85/90 kDa
Western blot - Human CTNNB1 knockout MCF7 cell line (ab286762)
False colour image of Western blot: Anti-beta Catenin antibody [SP328] staining at 1/400 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta Catenin antibody [SP328] ab224803 was shown to bind specifically to beta Catenin. A band was observed at 85/90 kDa in wild-type MCF7 cell lysates with no signal observed at this size in CTNNB1 knockout cell line ab286762. To generate this image, wild-type and CTNNB1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-beta Catenin antibody [SP328] (Anti-beta Catenin antibody [SP328] ab224803) at 1/400 dilution
Lane 1: Wild-type MCF7 cell lysate at 20 µg
Lane 2: CTNNB1 knockout MCF7 cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 85/90 kDa
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