Intracellular Staining by Flow Cytometry
|Detection of Smad3 in PC‑3 Human Cell Line by Flow Cytometry. PC‑3 human prostate cancer cell line was stained with Rat Anti-Human Smad3 Monoclonal Antibody (Catalog # MAB4038, filled histogram) or isotype control antibody (Catalog # MAB005, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG F(ab')2 Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.|
|Smad3 in Human Pancreatic Cancer Tissue. Smad3 was detected in immersion fixed paraffin-embedded sections of human pancreatic cancer tissue using 15 µg/mL Rat Anti-Human Smad3 Monoclonal Antibody (Catalog # MAB4038) 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-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
|Smad3 in MDA‑MB‑231 Human Cell Line. Smad3 was detected in immersion fixed MDA‑MB‑231 human breast cancer cell line using Rat Anti-Human Smad3 Monoclonal Antibody (Catalog # MAB4038) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (red; Catalog # NL013) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
Smad3 is phosphorylated in cells stimulated with TGF-beta, complexes with Smad4, and translocates to the nucleus to upregulate gene transcription. Smad3 is critical for signaling fibrosis and wound healing.
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