Murine and human carcinoma cells were treated for 72 h with 10 μM ARI-4175 and assessed for changes in a variety of immunologically relevant cell-surface molecules [17 (link)]. Murine cells were stained using the following antibodies: anti-H2Kb/H2Db-FITC, anti-H2Kd/H2Dd-APC, anti-CD54 (ICAM-1)-PE, anti-CD95 (Fas)-PE-Cy7 (BD Biosciences, San Diego, CA), anti-Col-1 (CEA)-FITC ([18 (link)]), and anti-Calreticulin-PE (R&D Systems, Minneapolis, MN). Human cells were assessed with: anti-CD95 (Fas)-PerCP-Cy5.5, anti-CD54 (ICAM-1)-PE, anti-CD227 (MUC1)-FITC, anti-HLA-A2-FITC, (BD Biosciences), anti-CD66 (CEA)-APC (Miltenyi Biotech, Auburn, CA) and anti-Calreticulin-PE (R&D Systems). Stained cells were acquired on an LSR II flow cytometer and analyzed using FloJo software. Isotype staining was < 5% for all samples analyzed. Proteins were scored as up-regulated if either the percent of cells or the mean fluorescence intensity (MFI) was increased by > 10% relative to untreated and vehicle-treated controls.
Anti cd227 muc1 fitc
Manufactured by BD
Anti-CD227 (MUC1)-FITC is a fluorescently labeled antibody that specifically binds to the CD227 (MUC1) antigen. It is designed for use in flow cytometry applications to detect and analyze cells expressing the CD227 (MUC1) marker.
Automatically generated - may contain errors
2 protocols using anti cd227 muc1 fitc
1
Immunological Changes in Carcinoma Cells Treated with ARI-4175
2
Immunophenotypic Changes in Tumor Cells
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Ovarian (OV17-1) and breast (MDA-MB-231) tumor cells were treated with the Cmax of mL4-3 and L1-7(N) or control (human IgG1-Fc) for 3 days. Cells were then harvested and analyzed for changes in expression of a number of cell-surface markers that have been reported to be immunologically relevant [34 (link)–38 (link)]. Cells were stained for 30 min at 4 °C using the following antibodies: anti-CD54/ICAM-1-PE, anti-CD95/Fas-FITC, anti-CD227/MUC-1-FITC, anti-HLA-A2-PerCP-Cy5.5 (BD Biosciences), anti-Trail-R1-PerCP, anti-Trail-R2-AF700, anti-calreticulin-PE (R&D Systems), and anti-CEA-APC (Miltenyi Biotec, San Diego, CA); appropriate isotype controls were used. Live/dead-Pacific Blue (Life Technologies) was included to discriminate viable cells. Stained cells were acquired with an LSRII flow cytometer (BD Biosciences) and analyzed using FlowJo software (TreeStar, Inc.). Isotype staining was < 5 % for all samples analyzed. Proteins were defined as upregulated by treatment with the angiopoietin inhibitors if either the percentage of cells or the MFI increased by > 10 % relative to cells treated with the IgG1-Fc control.
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