AB269661

Human TPBG knockout MCF7 cell lysate

Name: Human TPBG knockout MCF7 cell lysate (ab269661) | Abcam
Brand: Abcam Limited
SKU: AB269661
Price: 690 USD
Availability: InStock

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TPBG KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 97%.

View Alternative Names

5T4 oncofetal antigen, 5T4 oncofetal trophoblast glycoprotein, 5T4 oncotrophoblast glycoprotein, 5T4AG, AW495680, M6P1, TPBG_HUMAN, Trophoblast glycoprotein

4 Images

Lab

Western blot - Human TPBG knockout MCF7 cell lysate (AB269661)

Lane 1 : Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 2 : TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate, 20 ug
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lane 4 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab134162 observed at 85 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab134162 was shown to react with 5T4 in wild-type MCF7 cells in Western blot with loss of signal observed in TPBG knockout cell line ab269499 (knockout cell lysate ab269661). Wild-type MCF7 and TPBG knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in before incubation with ab134162 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-5T4 antibody [EPR5529] (<a href='/en-us/products/primary-antibodies/5t4-antibody-epr5529-ab134162'>ab134162</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TPBG knockout MCF7 cell line (<a href='/en-us/products/cell-lines/human-tpbg-knockout-mcf7-cell-line-ab269499'>ab269499</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 85 kDa

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Lab

Western blot - Human TPBG knockout MCF7 cell lysate (AB269661)

ab199547 was shown to react with 5T4 (HRP) in wild-type MCF7 cells in western blot. Loss of signal was observed when TPBG knockout cell line ab269499 (knockout cell lysate ab269661) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab199547 overnight at 4 °C at a 1 in 5000 dilution Blots were developed with Optiblot ECL reagent (ab133456) and imaged.

All lanes:

Western blot - HRP Anti-5T4 antibody [EPR5530] (<a href='/en-us/products/primary-antibodies/hrp-5t4-antibody-epr5530-ab199547'>ab199547</a>) at 1/5000 dilution

Lane 1:

Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TPBG knockout MCF7 cell line (<a href='/en-us/products/cell-lines/human-tpbg-knockout-mcf7-cell-line-ab269499'>ab269499</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 85 kDa

false

Lab

Western blot - Human TPBG knockout MCF7 cell lysate (AB269661)

Lanes 1 - 4 : Merged signal (red and green). Green - ab129058 observed at 85 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab129058 was shown to react with 5T4 in wild-type MCF7 cells in Western blot with loss of signal observed in TPBG knockout cell line ab269499 (knockout cell lysate ab269661). Wild-type MCF7 and TPBG knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in before incubation with ab129058 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-5T4 antibody [EPR5530] (<a href='/en-us/products/primary-antibodies/5t4-antibody-epr5530-ab129058'>ab129058</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TPBG knockout MCF7 cell line (<a href='/en-us/products/cell-lines/human-tpbg-knockout-mcf7-cell-line-ab269499'>ab269499</a>)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Predicted band size: 46 kDa

Observed band size: 85 kDa

false

Supplier Data

Next Generation Sequencing - Human TPBG knockout MCF7 cell lysate (AB269661)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 97%

Key facts

Cell type

MCF7

Species or organism

Human

Tissue

Breast

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 97%

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, 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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab269661-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human TPBG knockout MCF7 cell lysate", "number":"AB269661-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type MCF7 cell lysate", "number":"AB269661-CMP02", "productcode":"" } ] } } }

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Properties and storage information

Gene name

TPBG

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing, Western blot

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.

The 5T4 protein also known as trophoblast glycoprotein or B3F1 is a cell surface antigen with a mass of approximately 72 kDa. It is encoded by the TPBG gene and expresses mainly in embryonic tissues and various cancers with limited expression in normal adult tissues. The 5T4 protein is characterized by a densely glycosylated structure contributing to its role in cellular processes like adhesion and migration.

Biological function summary

5T4 is involved in processes associated with embryonic development and cellular mobility. It contributes to the epithelial-to-mesenchymal transition (EMT) which is a critical process in both normal development and cancer metastasis. Although 5T4 is not part of a known specific complex its glycoprotein nature allows interaction with the extracellular matrix and other cell surface proteins influencing cell behavior and morphology.

Pathways

5T4 plays a part in signaling pathways involved in cell migration and invasion such as the Wnt/β-catenin pathway and TGF-β signaling. In the context of the Wnt/β-catenin pathway 5T4 interacts with proteins like β-catenin influencing cell proliferation and survival. Through its involvement in TGF-β signaling 5T4 can impact cell differentiation processes and regulatory mechanisms often linking to other proteins such as SMAD family members within these pathways.

5T4 is associated with cancer particularly in its role in promoting metastasis. Its expression is notably increased in several carcinomas including breast and colorectal cancers. The elevated levels of 5T4 in tumors correlate with poorer prognosis making it a potential target for cancer therapies. Additionally 5T4 has connections to proteins such as MMPs (matrix metalloproteinases) which facilitate tissue remodeling and metastatic spread highlighting its significance in cancer progression.

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Cell culture

Biosafety level

EU: 1 US: 1

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 TPBG knockout MCF7 cell lysate

Western blot - Human TPBG knockout MCF7 cell lysate (AB269661)

Western blot - Human TPBG knockout MCF7 cell lysate (AB269661)

Western blot - Human TPBG knockout MCF7 cell lysate (AB269661)

Next Generation Sequencing - Human TPBG knockout MCF7 cell lysate (AB269661)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Complementary Products

Proteins & Peptides

Primary Antibodies

Cell Lines & Lysates

Assay Kits

Product promise