AB263756

Human CTNNB1 (beta Catenin) knockout HeLa cell lysate

Name: Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (ab263756) | Abcam
Brand: Abcam Limited
SKU: AB263756

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CTNNB1 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3.

3 Images

Lab

Western blot - Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (AB263756)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBST

The molecular weight observed is consistent with what has been described in the literature (PMID : 16288032).

Lane 1 : Wild-type HeLa cell lysate (20μg)**
**Lane 2 :****CTNNB1 knockout HeLa cell lysate (20μg)

Lanes 1-2 : Merged signal (red and green). Green - ab246504 observed at 92 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

ab246504 Anti-beta Catenin (non-phospho S37/T41) antibody [EPR23969-131] was shown to specifically react with beta Catenin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255352 (knockout cell lysate ab263756) was used.

Wild-type and beta Catenin knockout samples were subjected to SDS-PAGE. ab246504 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution 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 10000 dilution for 1 hour at room temperature before imaging.

An extra band around 45 kDa was observed.

All lanes:

Western blot - Anti-beta Catenin non-phospho S37/T41 antibody [EPR23969-131] (<a href='/en-us/products/primary-antibodies/beta-catenin-non-phospho-s37-t41-antibody-epr23969-131-ab246504'>ab246504</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

CTNNB1 (beta Catenin) knockout HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CTNNB1 (beta Catenin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ctnnb1-beta-catenin-knockout-hela-cell-line-ab255352'>ab255352</a>)

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 85 kDa

Observed band size: 95 kDa

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Lab

Western blot - Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (AB263756)

Lane 1 : Wild-type HeLa cell lysate (20µg)

Lane 2 : CTNNB1 knockout HeLa cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab32572 observed at 86 kDa. Red - loading control ab8245 observed at 37 kDa.

ab32572 Recombinant Anti-beta Catenin antibody [E247] - ChIP Grade was shown to specifically react with CTNNB1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255352 (knockout cell lysate ab263756) was used. Wild-type and CTNNB1 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32572 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4^°C at 1 in 500 dilution 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-beta Catenin antibody [E247] - ChIP Grade (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-e247-chip-grade-ab32572'>ab32572</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CTNNB1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CTNNB1 (beta Catenin) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ctnnb1-beta-catenin-knockout-hela-cell-line-ab255352'>ab255352</a>)

Predicted band size: 85 kDa

Observed band size: 86 kDa

false

Sanger Sequencing - Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (AB263756)

Sanger seq

Unknown

Sanger Sequencing - Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (AB263756)

Homozygous : 1 bp deletion in exon 3

Key facts

Cell type

HeLa

Species or organism

Human

Tissue

Cervix

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 3.

Disease

Adenocarcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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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 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.

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What's included?

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Properties and storage information

Gene name

CTNNB1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Shipped at conditions

Ambient - Can Ship with Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Human CTNNB1 (beta Catenin) knockout HeLa cell lysate

Western blot - Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (AB263756)

Western blot - Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (AB263756)

Sanger Sequencing - Human CTNNB1 (beta Catenin) knockout HeLa cell lysate (AB263756)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Complementary Products

Cell Lines & Lysates

Primary Antibodies

Secondary Antibodies

Proteins & Peptides

Reactivity data

Product details

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

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Product promise