Anti-5T4 antibody [EPR5529] (ab134162)

Anti-5T4 antibody [EPR5529] (ab134162) is a rabbit monoclonal antibody that is used to detect 5T4 in Western Blot, IHC-P. Suitable for Human samples.

- Specificity confirmed with 5T4 knockout cell line validation

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Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] (AB134162), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	Flow Cyt	WB	ICC/IF	IHC-P
Human	Not recommended	Not recommended	Tested	Not recommended	Tested

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000 - 1/10000	Notes For unpurified use at 1/500.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes For unpurified use at 1/100. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information TPBG

Function

May function as an inhibitor of Wnt/beta-catenin signaling by indirectly interacting with LRP6 and blocking Wnt3a-dependent LRP6 internalization.

Alternative names

5T4, TPBG, Trophoblast glycoprotein, 5T4 oncofetal antigen, 5T4 oncofetal trophoblast glycoprotein, M6P1, Wnt-activated inhibitory factor 1, 5T4 oncotrophoblast glycoprotein, WAIF1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR5529
Purification technique: Affinity purification Protein A
Dissociation constant: 1.5 x 10^-12 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-5T4 antibody [EPR5529] (ab134162) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunohistochemistry (IHC-P), in Human samples.

What is the molecular weight of 5T4?

Anti-5T4 [EPR5529] (ab134162) specifically detects a band for 5T4 (UniProt: Q13641) at a molecular weight of 46kDa.

Recommended positive controls

WB: MCF7, HeLa, HEK-293T, HT-29, HT-1376, and Caco-2 cell lysates.IHC-P: Human placenta and endometrial cancer tissues.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies. Specificity confirmed

The specificity of Anti-5T4 antibody [EPR5529] (ab134162) has been confirmed by Western blot testing in TPBG Knockout MCF7 cell line, ab269499.

Other related products

We have a range of other formats of antibody clone [EPR5529] also available for your convenience:
ab134162, Carrier free - ab201711

Species reactivity
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species.
Please contact us for more information.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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9 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] (ab134162)
Immunohistochemical staining using ab134162 on paraffin embedded human placenta tissue.
Western blot - Anti-5T4 antibody [EPR5529] (ab134162)
Lanes 1 - 4: Merged signal (red and green). Green - ab134162 observed at 85 kDa. Red - loading control Anti-GAPDH antibody [6C5] - 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 Human TPBG knockout MCF7 cell line ab269499 (knockout cell lysate Human TPBG knockout MCF7 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 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] (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
Western blot - Anti-5T4 antibody [EPR5529] (ab134162)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-5T4 antibody [EPR5529] (ab134162) at 1/500 dilution
All lanes: HT-29 cell lysate at 20 µg
Secondary
All lanes: Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 72-80 kDa
Western blot - Anti-5T4 antibody [EPR5529] (ab134162)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-5T4 antibody [EPR5529] (ab134162) at 1/5000 dilution
All lanes: HT-29 cell lysate at 20 µg
Secondary
All lanes: Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 46 kDa
Observed band size: 72-80 kDa
Western blot - Anti-5T4 antibody [EPR5529] (ab134162)
All lanes: Western blot - Anti-5T4 antibody [EPR5529] (ab134162) at 1/1000 dilution
Lane 1: HT-29 cell lysates at 10 µg
Lane 2: HT-1376 cell lysates at 10 µg
Lane 3: Caco-2 cell lysates at 10 µg
Secondary
All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size: 46 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] (ab134162)
Immunohistochemical staining using ab134162 on paraffin embedded human endometrial cancer.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] (ab134162)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labelling 5T4 with unpurified ab134162 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] (ab134162)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labelling 5T4 with purified ab134162 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
OI-RD Scanning - Anti-5T4 antibody [EPR5529] (ab134162)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.