Detection of Human Palladin by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, 293T human embryonic kidney cell line, PC‑3 human prostate cancer cell line, and Caki‑2 human clear cell carcinoma epithelial cell line. PVDF Membrane was probed with 0.5 µg/mL of Human Palladin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5950) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Palladin at approximately 95, 115, and 140 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Palladin in Human Prostate.
Palladin was detected in immersion fixed paraffin-embedded sections of human prostate using Human Palladin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5950) at 10 µg/mL 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 using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and nuclei of epithelial cells. 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 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
Palladin (a cytoarchitectural protein named after the 16th century Venetian architect Palladio) is a cytoplasmic member of the myotilin/palladin family of proteins. Palladin is widely expressed and interacts with actin-associated proteins, thus promoting actin bundle stability. Human Palladin is 1383 amino acids (aa) in length and contains five I-type Ig-like domains (aa 271-360; 440-539; 1001-1085; 1135-1226; 1233-1324), a Pro-rich region (aa 634-864) and nine potential Ser/Thr phosphorylation sites. Multiple isoform variants exist. Predominant are a 140 kDa and 92-95 kDa short form that show alternative start sites at Met383 and Met712, respectively. The 140 kDa isoform also shows a deletion of aa 656-879 to generate an approximately 115 kDa isoform.
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