Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate
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(0 Publication)
- WB
Lab
Western blot - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (AB275247)
Lane 1 : Wild-type HCT 116 cell lysate 20 μg
Lane 2 : CTNNB1 CRISPR-Cas9 edited 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 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 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:
CTNNB1 CRISPR-Cas9 edited HCT 116 cell lysate at 20 µg
Predicted band size: 85 kDa
Observed band size: 95 kDa
false
- WB
Lab
Western blot - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (AB275247)
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 (ab275247)
Lane 2:
Western blot - Human CTNNB1 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-ctnnb1-knockout-hct116-cell-line-ab273712'>ab273712</a>)
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 (beta Catenin) knockout HCT116 cell lysate (AB275247)
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 (ab275247)
Lane 2:
Western blot - Human CTNNB1 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-ctnnb1-knockout-hct116-cell-line-ab273712'>ab273712</a>)
Lane 2:
CTNNB1 knockout HCT116 cell lysate at 20 µg
Predicted band size: 85 kDa
Observed band size: 95 kDa
false
- WB
Lab
Western blot - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (AB275247)
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 [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 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 5 % 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 [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 knockout HCT 116 cell lysate at 20 µg
Predicted band size: 85 kDa
Observed band size: 95 kDa
false
- WB
Lab
Western blot - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (AB275247)
Lane 1 : Wild-type HCT 116 cell lysate 20 μg
Lane 2 : CTNNB1 CRISPR-Cas9 edited HCT 116 cell lysate 20 μg
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 °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 [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 (<a href='/en-us/products/cell-lines/human-ctnnb1-knockout-hct116-cell-line-ab273712'>ab273712</a>)
Predicted band size: 85 kDa
Observed band size: 95 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (AB275247)
Allele-1 : 7 bp insertion and 14 bp deletion in exon 3.
Complementary Products
Cell Lines & Lysates
AB277911
Human CTNNB1 (beta Catenin) knockout Hep G2 cell line
Advanced Validation
- 0
- 0
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)
- 5
- 75
Reactivity data
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.
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
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
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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