Description
• Uses traditional 3D soft agar matrix
• Fully quantify cell transformation with no manual cell counting
• Results in 7-8 days, not 3 weeks
Our CytoSelect 96-Well Cell Transformation Assay (Soft Agar Colony Formation) is suitable for measuring cell transformation where no downstream analysis is required. Cells are incubated in a semisolid agar medium for 7-8 days. The cells are then solubilized, lysed and detected using the included fluorescent dye in a fluorometric plate reader.
Introduction: Neoplastic transformation occurs via a series of genetic and epigenetic alterations that yield a cell population that is capable of proliferating independently of both external and internal signals that normally restrain growth. For example, transformed cells show reduced requirements for extracellular growth promoting factors, are not restricted by cell-cell contact, and are often immortal. Anchorage-independent growth is one of the hallmarks of transformation, which is considered the most accurate and stringent in vitro assay for detecting malignant transformation of cells.
Traditionally, the soft agar colony formation assay is a common method to monitor anchorage-independent growth, which measures proliferation in a semisolid culture media after 3-4 weeks by manual counting of colonies. Standard soft agar assays are usually performed in 100-mm or 60 mm dishes, where cells are allowed to grow inside a semisolid culture media for 3-4 weeks before sizable colonies appear. This method is quite cumbersome, time-consuming, and difficult when testing a large number of samples. Additionally, the manual counting of colonies is highly subjective; with varying colony sizes, it's difficult to determine meaningful results.
MyBioSource's CytoSelect 96-well Cell Transformation Assay 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 semisolid agar media before being solubilized, lysed and detected by the patented CyQuant GR Dye in a fluorescence plate reader (see Assay Principle below). This format provides a quantitative, high-throughput method to accurately measure cell transformation. Additionally, the short incubation time (6-8 days) makes it possible to assay cells transiently transfected with oncogenes or siRNA.
The CytoSelect 96-well Cell Transformation Kit provides a robust system for screening oncogenes and cell transformation inhibitors. Each kit provides sufficient quantities to perform 96 tests in a microtiter plate