Product nameStem Cell Colony Formation Assay Kit
The Stem Cell Colony Formation Assay Kit (ab242289) provides a stringent, anchorage- independent model for measuring stem cell colony formation and allows for cell recovery. Each kit provides sufficient quantities to perform 96 tests in a microtiter plate. The assay may be modified for use in other plate formats.
This kit does not involve subjective manual counting of colonies or require a 3-4 week incubation period. Instead cells are incubated only 6-8 days in a proprietary semisolid agar media before being solubilized, lysed and detected by the GR Dye in a fluorescence plate reader or assayed for alkaline phosphatase activity colorimetrically.
Alternatively, viable colony-forming cells (CFCs) can be easily recovered for further culturing and testing. This format provides a quantitative, high-throughput method to accurately measure colony formation, while the short incubation time makes it possible to assay transiently transfected cells.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Agar Matrix Solution 1 x 10ml 10X Matrix Stabilization Solution 1 x 1.8ml 5X DMEM Solution 3 x 1.5ml AP Activity Assay Standard 1 x 1ml AP Activity Assay Substrate 1 x 5ml AP Stop Solution 1 x 20ml GR Dye 1 x 75µl Lysis Buffer 1 x 20ml Matrix Diluent 1 x 4ml
Murine embryonic stem cells (ES-D3) were resuspended at 12 x 106 cells/mL and titrated 1:2 in culture medium, followed by addition of Cell Dose Curve Solution, Matrix Solubilization Solution, Lysis Buffer, and GR Dye detection (as described in the Cell Dose Section). Results are shown by actual cell number in dye Detection.
Murine embryonic stem cells (ES-D3) were cultured for 7 days in the presence (D-F) or absence (A-C) of LIF, according to the assay protocol. Phase contrast images demonstrate colonies at 10X magnification (A, D), 20X magnification (B, E), and 20X magnification with AP staining (C, F).
ab242289 has not yet been referenced specifically in any publications.