Quantibrite pe phycoreythrin fluorescence quantitation kit

Anti-human mesothelin antibody (Clone MB, Catalog No. 530203, BioLegend; San Diego, CA, USA) was conjugated with PE/R-phycoerythrin conjugation kit (Abcam, Cambridge, MA, USA; Catalog No. ab102918) per the manufacturer’s instructions. Samples stained with the PE-conjugated anti-MSLN antibody were analyzed on a NovoCyte 3000 Flow Cytometer. MSLN cell surface expression was quantitated using a BD Quantibrite PE Phycoreythrin Fluorescence Quantitation kit (BD Biosciences) following the manufacturer’s protocol.

CD123 expression was detected using a PE-conjugated anti-human CD123 antibody (Clone 6H6, Catalog No. 306006, BioLegend; San Diego, CA, USA). Samples were analyzed on a NovoCyte 3000 Flow Cytometer. CD123 cell surface expression was quantitated with the use of BD Quantibrite PE Phycoreythrin Fluorescence Quantitation kit (Catalog No. 340495, BD Biosciences, San Jose, CA, USA) following the manufacturer’s protocol.
Peripheral blood was stained with APC-conjugated anti-mouse CD45 (BioLegend Catalog No. 103112), Pacific blue conjugated anti-human CD45 (BioLegend Catalog No. 304029) and FITC conjugated anti-human CD3 (BioLegend Catalog No. 317306) antibodies following incubation with human BD Fc block (BD Biosciences, Catalog No. 564219) to prevent non-specific antibody binding.