Monitor Stem Cell Differentiation and Pathway Activation Using Multi-color FlowCellect Analysis on Guava Bench Top Flow Cytometer
Matthew Hsu, Ph.D.
R&D Director, Millipore USA
Flow cytometry provides high quality, reproducible data for cellular analysis in far less time than traditional methods. The ability to simultaneously measure multiple parameters on a cell by cell basis is probably the most powerful aspect of flow cytometry. Scientists consider multi-parameter flow cytometry analysis as a better way to monitor phenotypic change and pathway activation, as they get multiple readouts and confirmation on a particular cellular event within different cell populations.
We have developed a series of cell-based assays for monitoring stem cell differentiation, GPCR and kinase pathway activation using multi-color FlowCellectTM Analysis. FlowCellectTM are a group of fully optimized multi-parameter flow cytometry kits that we developed in house, which contain directly conjugated and validated Abs and Ab-compatible buffers for cellular event analysis and possibly cell sorting （cell selection）， they are “plug and play” solutions for scientists to study cell biology events with speed and accuracy by monitoring the changes of protein expression and post-translational modification in individual cells.
Guava EasyCyte? bench top flow cytometer and its microcapillary “flow cell” technology enable the analysis of very few cells, thus making the platform ideally suited to study stem cell differentiation. The FlowCellect? stem cell kits that can be used to detect and quantify changes （or lack of changes） of protein markers in both embryonic and adult stem cell populations, FlowCellect? stem cell kits provide a faster and reliable alternative to obtaining multiple endpoint （phenotypic） information on a mix cell population and track the stem cell differentiation.
The FlowCellect? phospho detection kits provide powerful research tools for measuring intracellular kinase activity by delivering robust, high content information on a cell-by-cell basis to measure multiple parallel or vertical end points of an upstream kinase activity in spatial and temporal manner. Novel GPCR 2nd assays to monitor receptor internalization and GPCR-mediated ERK phosphorylation will be presented. The receptor internalization assay uses flow-validated GPCR Abs to selectively quantify the GPCR expression level only on the cell surface in 96 well format, thus allow monitoring the changes of these surface receptor expression levels as an indication of receptor internalization. The EC50 and IC50 of the agonists and antagonists correlate very well with the FLIPR assay.
编辑: ludongcn 作者:Matthew Hsu, Ph.D.
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