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%.
Images
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%
Alternative names
5T4 oncofetal antigen, 5T4 oncofetal trophoblast glycoprotein, 5T4 oncotrophoblast glycoprotein, 5T4AG, AW495680, M6P1, TPBG_HUMAN, Trophoblast glycoprotein
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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%.
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
- Concentration
Properties
- Gene name
- TPBG
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing, Western blot
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
Storage
- Shipped at conditions
- Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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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4 product images
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 ugLanes 1 - 4: Merged signal (red and green). Green - Anti-5T4 antibody [EPR5530] ab129058 observed at 85 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
Anti-5T4 antibody [EPR5530] 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 Human TPBG knockout MCF7 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 Anti-5T4 antibody [EPR5530] ab129058 and Anti-GAPDH antibody [6C5] - Loading Control 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 (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) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-5T4 antibody [EPR5530] (Anti-5T4 antibody [EPR5530] ab129058) 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 (Human TPBG knockout MCF7 cell line ab269499)
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
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 85 kDa
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 ugLanes 1 - 4: Merged signal (red and green). Green - Anti-5T4 antibody [EPR5529] ab134162 observed at 85 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.
Anti-5T4 antibody [EPR5529] 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 Human TPBG knockout MCF7 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 Anti-5T4 antibody [EPR5529] ab134162 and Anti-GAPDH antibody [6C5] - Loading Control 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 (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) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
All lanes: Western blot - Anti-5T4 antibody [EPR5529] (Anti-5T4 antibody [EPR5529] ab134162) 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 (Human TPBG knockout MCF7 cell line ab269499)
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
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 85 kDa
Next Generation Sequencing - Human TPBG knockout MCF7 cell lysate (ab269661)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 97%
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 ugHRP Anti-5T4 antibody [EPR5530] 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 Human TPBG knockout MCF7 cell line ab269499 (knockout cell lysate ab269661) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with HRP Anti-5T4 antibody [EPR5530] ab199547 overnight at 4 °C at a 1 in 5000 dilution Blots were developed with Optiblot ECL reagent (Anti-PKC delta (phospho S299) antibody [EPNCI119] ab133456) and imaged.
All lanes: Western blot - HRP Anti-5T4 antibody [EPR5530] (HRP Anti-5T4 antibody [EPR5530] ab199547) 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 (Human TPBG knockout MCF7 cell line ab269499)
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
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 85 kDa
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