RT-PCR Measurement of each Marker :The secretory protein has a good correlation with Sox 17 and Foxa2 which are commonly used proteins as markers of endoderm differentiation.
Comparison with Conventional Techniques :
| Dojindo Kit | PCR | Immunostaining | Flow Cytometry | |
| Procedure | 3 hours | > 7 hours | 1-2 days | > 4 hours |
| Sample | Supernatant | mRNA | Whole cells | Whole cells |
| ContinuousCell Culture | Possible | Impossible | Impossible | Impossible |
| Marker | Marker Protein | Sox17, Foxa2 | Sox17, Foxa2 | CXCR4 |
Measurement of Procedure :
mmunocytochemical(ICC) Analysis:[Sample: Murine ES cell]
Most of the Foxa2 positive cells are co-stained with the marker protein.The three images in the upper row were obtained from conventional ICC analysis. Normally, Foxa2 and Sox17 double positive cells are defined as differentiated into endodermal cells.
Differentiation Monitoring by ICC and ELISA:[Sample: Human iPS cell]
Human iPS 253G1 cells were differentiated into the definitive endoderm. They were being monitored with ELISA assay to measure the amount of marker protein secreted or ICC analysis for Sox17 and Foxa2 double positive cells.
The two bar graphs show the correlation of the amount of secreted protein with the amount of Sox17 and Foxa2 double positive cells from day 1 to day 5 (D1 to D5) of differentiation.