Detection of Human 5T4 by Western Blot.
Western blot shows lysates of HT‑29 human colon adenocarcinoma cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human 5T4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4975) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for 5T4 at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
5T4 in Human Placenta.
5T4 was detected in immersion fixed paraffin-embedded sections of human placenta using 10 µg/mL Sheep Anti-Human 5T4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4975) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent‑Basic (Catalog # CTS013). Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
5T4 in Human Placenta.
5T4 was detected in immersion fixed paraffin-embedded sections of human placenta using Sheep Anti-Human 5T4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4975) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Human 5T4 (oncofetal antigen 5T4; also TPBG and trophoblast glycoprotein) is a 72 kDa glycoprotein member of the LRR family of proteins. It is expressed on trophoblasts, tumor cells, ovarian cuboidal epithelium, and embryonic stem cells and impacts cell adhesion and motility. The human 5T4 cDNA encodes a type I transmembrane protein precursor that is 420 amino acids (aa) in length. It contains a 324 aa extracellular region (aa 32-355) that shows one Ser-rich region followed by seven Leu-rich repeats (aa 90-355). Over aa 31-355, human 5T4 shares 81% and 85% aa identity with mouse and canine 5T4, respectively.
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