Separation of MCF-7 cells (red-filled) from human leukocytes (black-dashed) in flow cytometry analysis (intracellular staining) of peripheral whole blood spiked with MCF-7 cells stained using anti-Cytokeratins (C-11) purified antibody (concentration in sample 3 µg/ml, GAM APC).
Flow cytometry intracellular staining pattern of human peripheral whole blood spiked with MCF-7 cells stained using anti-Cytokeratins (C-11) purified antibody (concentration in sample 3 µg/ml, GAM APC).
Immunocytochemistry staining of cytokeratins in Hep-2 cells using pan-cytokeratin antibody C-11 (orb43707, diluted 1:400), detected with GAM IgG-Alexa Fluor488 (diluted 1:200, green).
Immunohistochemistry staining of cytokeratin on paraffin-embedded sections of guinea pig breast carcinoma using anti-cytokeratin antibody (C-11).
Western blotting analysis of human cytokeratins using mouse monoclonal antibody C-11 on lysates of HeLa cell line and MOLT-4 cell line (cytokeratin non-expressing cell line, negative control) under non-reducing and reducing conditions. Nitrocellulose membrane was probed with 2 µg/ml of mouse monoclonal antibody anti-cytokeratins followed by IRDye800-conjugated anti-mouse secondary antibody. Specific bands were detected for cytokeratins at approximately 45-55 kDa.
Anti-Hu Cytokeratin 5/8 (clone C-50) works in WB application under reducing and non-reducing conditions. Western blotting analysis was performed on whole cell extracts (RIPA lysis buffer) of MCF-7 and Jurkat cell lines, mixed and heated (100C, 5 min) with reducing and non-reducing SDS-loading buffer. Samples were resolved using 10% Tris-glycine SDS gel electrophoresis. Nitrocellulose membrane blot was probed with mouse IgG1 monoclonal antibody C-50 (1 µg/ml), followed by IRDye 800CW Goat-anti-Mouse IgG (green). Mouse anti-GAPDH monoclonal antibody FF26A conjugated with DyLight 680 (0.1 µg/ml), was used as the loading control (red). Multiplex fluorescent Western blot detection was performed. Cytokeratin 5/8 molecules were detected at ~40 kDa in MCF-7 cell line under both reducing and non-reducing conditions.
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