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 (HRP)
- Suitable for: WB, IHC-P
- Reacts with: Human
- Conjugation: HRP
Product nameAnti-5T4 antibody [EPR5530] (HRP)
See all 5T4 primary antibodies
DescriptionRabbit monoclonal [EPR5530] to 5T4 (HRP)
Tested applicationsSuitable for: WB, IHC-Pmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
Synthetic peptide within Human 5T4 aa 350-450 (intracellular). The exact sequence is proprietary.
- WB: MCF7 whole cell lysate. IHC-P: FFPE human placenta (normal) tissue sections.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark.
Storage bufferpH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: PBS, 30% Glycerol, 1% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab199547 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000. Detects a band of approximately 72-80 kDa (predicted molecular weight: 46 kDa).|
|IHC-P||1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
Tissue specificityExpressed 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 similaritiesContains 6 LRR (leucine-rich) repeats.
Contains 1 LRRCT domain.
Contains 1 LRRNT domain.
- Information by UniProt
Form5T4 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).
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IHC image of 5T4 staining in a section of formalin-fixed paraffin-embedded human placenta (normal)*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab199547, 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Anti-5T4 antibody [EPR5530] (HRP) (ab199547) at 1/5000 dilution + MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 46 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?
Exposure time: 20 minutes
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab199547 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
ab199547 has not yet been referenced specifically in any publications.