CTNNB1 (beta Catenin II) KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion and 7 bp insertion in exon 3.
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
CTNNB1 (beta Catenin II) KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion and 7 bp insertion in exon 3.
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.
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.
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.
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.
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.
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.
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All lanes: Western blot - Anti-beta Catenin antibody [IGX4794R-3] (Anti-beta Catenin antibody [IGX4794R-3] ab223075) at 1 µg/mL
Lane 1: Wild-type HCT116 cell lysate at 20 µg
Lane 2: Western blot - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate ab275247)
Lane 2: Western blot - Human CTNNB1 knockout HCT116 cell line (ab273712)
Lane 2: CTNNB1 knockout HCT116 cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 95 kDa
False colour image of Western blot: Anti-beta Catenin antibody [E247] - ChIP Grade staining at 1/5000 dilution shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) 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 95 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CTNNB1 CRISPR-Cas9 edited cell line ab273712 (CRISPR-Cas9 edited cell lysate Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate ab275247). The band observed in the CRISPR-Cas9 edited lysate lane below 95 kDa is likely to represent a truncated form of beta Catenin. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image wild-type and CTNNB1 CRISPR-Cas9 edited HCT 116 cell lysates were analysed. First samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °. 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) 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 HCT 116 cell lysate at 20 µg
Lane 2: CTNNB1 CRISPR-Cas9 edited HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human CTNNB1 knockout HCT116 cell line (ab273712)
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 95 kDa
All lanes: Western blot - Anti-beta Catenin antibody [IGX4794R-3] (Anti-beta Catenin antibody [IGX4794R-3] ab223075) at 1 µg/mL
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Western blot - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate ab275247)
Lane 2: Western blot - Human CTNNB1 knockout HCT116 cell line (ab273712)
Lane 2: CTNNB1 knockout HCT 116 cell lysate at 20 µg
Lanes 1 - 2: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/20000 dilution
Lanes 1 - 2: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 85 kDa
Observed band size: 95 kDa
Allele-1: 7 bp insertion and 14 bp deletion in exon 3.
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