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 hla a2 fitc
Manufactured by BD
Anti-HLA-A2-FITC is a fluorescently labeled antibody that binds specifically to the HLA-A2 antigen. It can be used for the detection and analysis of cells expressing HLA-A2 by flow cytometry.
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Lab products found in correlation
Anti calreticulin pe, by R&D Systems (1 mentions)
Anti cd227 muc1 fitc, by BD (1 mentions)
Anti cd54 icam 1 pe, by BD (1 mentions)
Anti cd95 fas pe cy7, by BD (1 mentions)
Anti pd 1 pe, by BioLegend (1 mentions)
Anti ccr7 alexa fluor 647, by BD (1 mentions)
Fab32652p, by R&D Systems (1 mentions)
Goat anti human igg f ab 2 biotin, by Bio-Rad (1 mentions)
Streptavidin apc, by Bio-Rad (1 mentions)
Anti cd86 percp cy5, by BD (1 mentions)
Anti tim3 apc, by BioLegend (1 mentions)
Facs cantoi 2, by BD (1 mentions)
Anti cd8 bv421, by BD (1 mentions)
2 protocols using anti hla a2 fitc
1
Immunological Changes in Carcinoma Cells Treated with ARI-4175
2
Mesothelin Expression on Human T-cells
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CAR Mesothelin surface expression on human T-cells was assessed using primary antibody goat anti-human IgG F (ab’)2 Biotin (BioRad, Hercules, CA) along with secondary conjugate Streptavidin APC (BioRad). The following antibodies were also used in FACS buffer in this study: anti-PD1-PE (Biolegends 329906), anti-CCR7-Alexa-fluor 647 (BD 560921), anti-CD8-FITC (BC A07756), anti-CD8-BV421 (BD 562428), anti-HLA-A2-FITC (BD 551,285), anti-CD86-PerCp-Cy5.5 (BD 561129), anti-CLDN6-Dylight 650 (IMAB027), anti-idiotype-IMAB027- Alexa-fluor 647, anti-TIM3-APC (Biolegends 345011) and 7AAD (BC A07704). Mesothelin expression was detected using the PE conjugated anti mesothelin antibody (R&D system FAB32652P). Acquisition and analysis of all samples were performed on a BD FACS CantoI/II (BD Biosciences, San Jose, California) and FlowJo software (v7.6.1).
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