Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)

Rabbit Recombinant Monoclonal 5T4 antibody. Carrier free. Suitable for WB, IHC-P and reacts with Human samples.

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Images

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (AB201711), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
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 ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

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.

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

Recommended products

Rabbit Recombinant Monoclonal 5T4 antibody. Carrier free. Suitable for WB, IHC-P and reacts with Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
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 conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab201711 is the carrier-free version of Anti-5T4 antibody [EPR5529] ab134162.

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

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

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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:
Terms & Conditions.

9 product images

Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)
This data was developed using Anti-5T4 antibody [EPR5529] ab134162, the same antibody clone in a different buffer formulation:
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711) 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)
Immunohistochemical staining using Anti-5T4 antibody [EPR5529] ab134162 on paraffin embedded human placenta tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-5T4 antibody [EPR5529] ab134162).
Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)
This data was developed using the same antibody clone in a different buffer formulation (Anti-5T4 antibody [EPR5529] ab134162).
Lanes 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 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 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
Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)
This data was developed using Anti-5T4 antibody [EPR5529] ab134162, the same antibody clone in a different buffer formulation.
All lanes: Western blot - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711) 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] - BSA and Azide free (ab201711)
This data was developed using Anti-5T4 antibody [EPR5529] ab134162, the same antibody clone in a different buffer formulation
All lanes: Western blot - Anti-5T4 antibody [EPR5529] (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] - BSA and Azide free (ab201711)
Immunohistochemical staining using Anti-5T4 antibody [EPR5529] ab134162 on paraffin embedded human endometrial cancer.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-5T4 antibody [EPR5529] ab134162).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labelling 5T4 with unpurified Anti-5T4 antibody [EPR5529] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-5T4 antibody [EPR5529] ab134162).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human placenta tissue labelling 5T4 with purified Anti-5T4 antibody [EPR5529] 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-5T4 antibody [EPR5529] ab134162).
OI-RD Scanning - Anti-5T4 antibody [EPR5529] - BSA and Azide free (ab201711)
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.