Recombinant Anti-5T4 antibody [EPR5530] (ab129058)
![Western blot - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-459641-anti-5t4-antibody-epr5530-western-blot-mcf7-wt-mcf7-tpbg-knockout-hela-hek293t.jpg "Western blot - Anti-5T4 antibody [EPR5530] (ab129058)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5530] to 5T4
- Suitable for: WB, IP
- Knockout validated
- Reacts with: Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-5T4 antibody [EPR5530]
See all 5T4 primary antibodies -
Description
Rabbit monoclonal [EPR5530] to 5T4 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IPmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC-P -
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse
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Immunogen
Synthetic peptide within Human 5T4 aa 350-450 (intracellular). The exact sequence is proprietary.
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Positive control
- HEK-293T, HT 1376, HeLa, JAR and MCF7 cell lysates; MCF7 cells.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5530 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab129058 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
1/1000 - 1/10000. Detects a band of approximately 72-80 kDa (predicted molecular weight: 46 kDa).
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| IP |
1/80.
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| Notes |
|---|
|
WB
1/1000 - 1/10000. Detects a band of approximately 72-80 kDa (predicted molecular weight: 46 kDa). |
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IP
1/80. |
Target
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Tissue specificity
Expressed by all types of trophoblasts as early as 9 weeks of development. Specific for trophoblastic cells except for amniotic epithelium. In adult tissues, the expression is limited to a few epithelial cell types but is found on a variety of carcinoma. -
Sequence similarities
Contains 6 LRR (leucine-rich) repeats.
Contains 1 LRRCT domain.
Contains 1 LRRNT domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 7162 Human
- Entrez Gene: 21983 Mouse
- Entrez Gene: 83684 Rat
- Omim: 190920 Human
- SwissProt: Q13641 Human
- SwissProt: Q9Z0L0 Mouse
- SwissProt: Q5PQV5 Rat
- Unigene: 82128 Human
see all -
Form
5T4 is a 72 kDa transmembrane glycoprotein that exhibits restricted expression in human and mouse adult tissues, is upregulated on many carcinomas and tumour expression correlates with poorer clinical outcome in ovarian, gastric and colorectal cancers. mES cells lack cell surface expression of the 5T4 antigen, and both protein and mRNA are rapidly upregulated following induction of differentiation. This proteins expression correlates with downregulation of OCT-4 and Tra-1–60 (Ward et al., 2006). -
Alternative names
- 5T4 antibody
- 5T4 oncofetal antigen antibody
- 5T4 oncofetal trophoblast glycoprotein antibody
see all
Images
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Western blot - Anti-5T4 antibody [EPR5530] (ab129058)All lanes : Anti-5T4 antibody [EPR5530] (ab129058) at 1/1000 dilution
Lane 1 : Wild-type MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : TPBG knockout MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?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.
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![Western blot - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-237655-anti-5t4-antibody-epr5530-western-blot.jpg)
Western blot - Anti-5T4 antibody [EPR5530] (ab129058)Anti-5T4 antibody [EPR5530] (ab129058) at 1/5000 dilution (purified) + Rat brain at 20 µg
Secondary
HRP goat-anti rabbit (H+L) at 1/1000 dilution
Predicted band size: 46 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Western blot - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-237653-anti-5t4-antibody-epr5530-western-blot.jpg)
Western blot - Anti-5T4 antibody [EPR5530] (ab129058)Anti-5T4 antibody [EPR5530] (ab129058) at 1/2000 dilution (unpurified) + Rat brain at 20 µg
Secondary
HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 46 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Western blot - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-237652-anti-5t4-antibody-epr5530-western-blot.jpg)
Western blot - Anti-5T4 antibody [EPR5530] (ab129058)All lanes : Anti-5T4 antibody [EPR5530] (ab129058) at 1/5000 dilution (purified)
Lane 1 : HT-1376 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 46 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Western blot - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-237648-anti-5t4-antibody-epr5530-western-blot.jpg)
Western blot - Anti-5T4 antibody [EPR5530] (ab129058)All lanes : Anti-5T4 antibody [EPR5530] (ab129058) at 1/2000 dilution (unpurified)
Lane 1 : HT-1376 cell lysate
Lane 2 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 46 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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![Western blot - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-142983-anti-5t4-antibody-epr5530-western-blot.jpg)
Western blot - Anti-5T4 antibody [EPR5530] (ab129058)All lanes : Anti-5T4 antibody [EPR5530] (ab129058) at 1/1000 dilution (unpurified)
Lane 1 : HT 1376 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : JAR cell lysate
Lane 4 : MCF-7 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP conjugated at 1/2000 dilution
Predicted band size: 46 kDa -
![Immunoprecipitation - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-237656-anti-5t4-antibody-epr5530-immunoprecipitation.jpg)
Immunoprecipitation - Anti-5T4 antibody [EPR5530] (ab129058)ab129058 (unpurified) at 1/50 immunoprecipitating 5T4 in MCF-7 (Lane 1). For western blotting, a HRP goat anti-rabbit was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Immunoprecipitation - Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-237657-anti-5t4-antibody-epr5530-immunoprecipitation.jpg)
Immunoprecipitation - Anti-5T4 antibody [EPR5530] (ab129058)ab129058 (unpurified) at 1/50 immunoprecipitating 5T4 in MCF-7 (Lane 1). For western blotting, a HRP goat anti-rabbit was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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![Anti-5T4 antibody [EPR5530] (ab129058)](https://www.abcam.com/ps/products/129/ab129058/Images/ab129058-9-benefits-of-recombinant-antibodies.png)
Anti-5T4 antibody [EPR5530] (ab129058)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab129058 has been referenced in 4 publications.
- Wakeham CM et al. Expression and distribution of trophoblast glycoprotein in the mouse retina. J Comp Neurol 528:1660-1671 (2020). PubMed: 31891182
- Wakeham CM et al. Identification of PKCa-dependent phosphoproteins in mouse retina. J Proteomics 206:103423 (2019). PubMed: 31255707
- Puchert M et al. The 5T4 oncofetal glycoprotein does not act as a general organizer of the CXCL12 system in cancer cells. Exp Cell Res 364:175-183 (2018). PubMed: 29408206
- Shor B et al. Enhanced Antitumor Activity of an Anti-5T4 Antibody-Drug Conjugate in Combination with PI3K/mTOR inhibitors or Taxanes. Clin Cancer Res 22:383-94 (2016). WB ; Human . PubMed: 26319086