Human CTNNB1 knockout HCT116 cell line
- Advanced Validation
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(1 Publication)
CTNNB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion and 7 bp insertion in exon 3. 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
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
- WB
Lab
Western blot - Human CTNNB1 knockout HCT116 cell line (AB273712)
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] (ab8245) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-beta Catenin antibody [E247] - ChIP Grade (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-e247-chip-grade-ab32572'>ab32572</a>) 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)
Predicted band size: 85 kDa
Observed band size: 95 kDa
false
- WB
Lab
Western blot - Human CTNNB1 knockout HCT116 cell line (AB273712)
Lane 1 : Wild-type HCT 116 cell lysate 20 μg
Lane 2 : CTNNB1 knockout HCT 116 cell lysate 20 μg
False colour image of Western blot : Anti-beta Catenin antibody [IGX4794R-3] staining at 1 μg/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab223075 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 knockout cell line ab273712 (knockout cell lysate ab275247). The band observed in the knockout 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 knockout 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 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-beta Catenin antibody [IGX4794R-3] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-igx4794r-3-ab223075'>ab223075</a>) 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 (<a href='/en-us/products/cell-lysates/human-ctnnb1-beta-catenin-knockout-hct116-cell-lysate-ab275247'>ab275247</a>)
Lane 2:
Western blot - Human CTNNB1 knockout HCT116 cell line (ab273712)
Lane 2:
CTNNB1 knockout HCT 116 cell lysate at 20 µg
Secondary
Lanes 1 - 2:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution
Lanes 1 - 2:
Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution
Predicted band size: 85 kDa
Observed band size: 95 kDa
false
- WB
Lab
Western blot - Human CTNNB1 knockout HCT116 cell line (AB273712)
Lane 1 : Wild-type HCT116 cell lysate 20 ug
Lane 2 : CTNNB1 knockout HCT116 cell lysate 20 ug
Lanes 1 - 2 : Merged signal (red and green). Green - ab223075 observed at 95 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.
ab223075 was shown to react with Anti-beta Catenin in wild-type HCT 116 cells in western blot with loss of signal observed in CTNNB1 knockout cell line ab273712 (CTNNB1 knockout cell lysate ab275247). HCT 116 wild-type and CTNNB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluoroscent western blot (TBS-based) blocking solution 50% (v/v) in TBS-T (0.1% Tween®) before incubation with ab223075 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 ug/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-beta Catenin antibody [IGX4794R-3] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-igx4794r-3-ab223075'>ab223075</a>) 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 (<a href='/en-us/products/cell-lysates/human-ctnnb1-beta-catenin-knockout-hct116-cell-lysate-ab275247'>ab275247</a>)
Lane 2:
Western blot - Human CTNNB1 knockout HCT116 cell line (ab273712)
Lane 2:
CTNNB1 knockout HCT116 cell lysate at 20 µg
Predicted band size: 85 kDa
Observed band size: 95 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CTNNB1 knockout HCT116 cell line (AB273712)
Allele-1 : 7 bp insertion and 14 bp deletion in exon 3.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
Zygosity
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
McCoY5a + 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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Publications (1)
Recent publications for all applications. Explore the full list and refine your search
Cell death discovery 9:132 PubMed37076452
2023
Applications
Unspecified application
Species
Unspecified reactive species
Complementary Products
Primary Antibodies
AB32572
Anti-beta Catenin antibody [E247] - ChIP Grade
BOND RX™ Validated|Advanced Validation|RabMAb|Recombinant|KO Validated|Lab Essentials|20ul selling size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [E247] - ChIP Grade (AB32572)](https://content.abcam.com/products/images/beta-catenin-antibody-e247-chip-grade-ab32572--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img93749.jpg)
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