Miltenyi Biotec-tested panel 22 (MBTP 22)
This application protocol describes the quantitative analysis of human pluripotent stem cell (PSC) differentiation potential using flow cytometry. Human PSCs can easily be differentiated using the StemMACS™ Trilineage Differentiation Kit which enables directed and reproducible differentiation into ectodermal, mesodermal, and endodermal lineages ready for downstream applications, such as flow cytometric analysis.
The staining panel is based on a set of REAfinity™ Antibodies to ensure low background and high staining specificity. The panel consists of the three marker combinations (combination 1: CD144 and CD140b for mesoderm analysis; combination 2: CD184 and Sox17 for endoderm analysis; combination 3: PAX-6 and Sox2 for ectoderm analysis) and includes both surface and intracellular markers. Surface markers are stained first. Subsequently, cells are fixed and permeabilized for intracellular staining. Since the marker combination for ectoderm detection only includes intracellular markers, no surface staining is needed for this sample.
|Specificity||Clone||Purpose||Fluorochrome||Detection filter (nm) (laser)|
|CD144 (VE-Cadherin)||REA199||Mesoderm||FITC||525/50 (blue)|
|CD184 (CXCR4)||REA649||Endoderm||APC||655-730 (red)|
|Sox17||REA701||Endoderm||Vio® B515||525/50 (blue)|
|Propidium Iodide Solution||-||Viability||-||655-730 (blue)|
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