CytoSelect™ Cell Transformation Assays

Transformation of normal cells into neoplastic cells results in a population capable of proliferating independently of internal and external signals that normally restrain growth. The soft agar colony formation assay has traditionally been used to monitor ancorageindependent growth, employing 3-4 weeks of cell growth followed by manuel cell counting.

We have advanced the soft agar assay to eliminate tedious manual cell counting, allow high-throughput drug screening, and enable recovery of transformed cells for downstream analysis. These advances have also allowed us to develop a unique kit for the separation of clonogenic cancer cells from normal cells in heterogeneous solid tumors.

We offer a variety of assay formats:

  • Cell Biolabs CytoSelect™ 96-Well Cell Transformation Assay, Standard Soft Agar combines a soft agar medium with a fluorescent dye to eliminate manual cell counting and speed results.
  • The CytoSelect™ 96-Well Cell Transformation Assay, Soft Agar with Cell Recovery advances the technology even further. This assay uses a proprietary soft agar-based medium that allows transformed cells to be recovered after culture for further downstream analysis.
  • For even higher throughput, choose our CytoSelect™ 384-Well Cell Transformation Assay which uses the proprietary medium found in our Cell Recovery Compatible assay. Due to small well size, it is not recommended to use this format for recovery of transformed cells.

CytoSelect™ 96-Well Cell Transformation Assay-Traditional Soft Agar Colony Formation

The CytoSelect™ 96-Well Cell Transformation Assay (Soft Agar Colony Formation) is suitable for measuring malignant transformation where no downstream analysis is required. Transformed cells cannot be recovered; however, no manual cell counting is required.

With this assay, cells are incubated in a semisolid agar medium for 6-8 days, then solubilized, lysed and detected using CyQuant® GR dye in a fluorometric plate reader.
  • Fast Results: 6-8 days vs. 21 days
  • Plate Reader Convenience: Eliminates manual counting
  • Versatile Format: Designed for 96-well throughput, but van be adapted for 48, 24, 12 or 6-well
Cat.No.DescriptionAmount
CBA-130
CytoSelect 96-well Cell Transformation Assay
96 assays
CBA-130-5
CytoSelect 96-well Cell Transformation Assay
5 x 96 assays

Recent Product Citations

  1. Faoro, L. et al. (2010). EphA2 Mutation in Lung Squamous Cell Carcinoma Promotes Increased Cell Survival, Cell Invasion, Focal Adhesions, and Mammalian Target of Rapamycin Activation. J. Biol. Chem. 285:18575-18585.
  2. Carnahan, J. (2010). Selective and Potent Raf Inhibitors Paradoxically Stimulate Normal Cell Proliferation and Tumor Growth. Mol. Cancer Ther. 9:2399-2410.
  3. Liu, F. et al. (2010). Epigenomic alterations and gene expression profiles in respiratory epithelia exposed to cigarette smoke condensate. Oncogene 29:3650-3664.
  4. Iorns, E. et al. (2010). The Role of SATB1 in Breast Cancer Pathogenesis. J. Natl. Cancer Inst. 10.1093/jnci/djq243.
  5. Rubio, R. et al. (2010). Deficiency in p53 but not retinoblastoma induces the transformation of mesynchymal stem cells in vitro and initiates leiomyosarcoma in vivo. Cancer Res. 70:4185-4194.
  6. Takezawa, K. et al. (2009). Sorafenib inhibits non-small cell lung cancer cell growth by targeting B-RAF in KRAS wild-type cells and C-RAF in KRAS mutant cells. Cancer Res. 69:6515-6521.
  7. Li, H. et al. (2009). Lysophosphatidic acid stimulates cell migration, invasion, and colony formation as well as tumorigenesis/metastasis of mouse ovarian cancer in immunocompetent mice. Mol. Cancer Ther. 8:1692-1701.
  8. Kang, M-I. et al. (2009). A selective small-molecule nuclear factor-kB inhibitor from a high-throughput cell-based assay for "activator protein-1 hits". Mol. Cancer Ther. 8:571-581.
  9. Lee, K.B. et al. (2008). Low energy proton beam induces tumor cell apoptosis through reactive oxygen species and activation of caspases. Exp. and Mol. Medicine 40(1):118-129.
  10. Shen, L. et al. (2008). E1A inhibits the proliferation of human cervical cancer cells (HeLa cells) by apoptosis induction through activation of HER-2/Neu/Caspase-3 pathway. Med. Oncol. 25:222-228.
  11. Wei, Q. et al. (2008). Sulfiredoxin is an AP-1 target gene that is required for transformation and shows elevated expression in human skin malignancies. PNAS 105:19738-19743.
  12. Gazin, C. et al. (2007). An elaborate pathway required for Ras-mediated epigenetic silencing. Nature 449(7165):1073-1077.

CytoSelect™ 96-Well Cell Transformation Assays-Advanced Soft Agar with Post-Incubation Cell Recovery

The CytoSelect™ 96-Well Cell Transformation Assay (Cell Recovery Compatible) provides a robust system for screening oncogenes and cell transformation inhibitors. Transformed cells may be recovered for further downstream analysis following colony formation.
  • Faster Results: 6-8 days vs. 21 days
  • Cell Recovery: Transformed cells remain viable for further analysis
  • Plate Reader Convenience: Eliminates manual counting of cells
  • Versatile Format: Designed for 96-Well through-put, but can be adapted for 48, 24, 12 or 6-well

Picture: Cell Transformation Assay Principle. Cell colonies form after a 6-8 day incubation with agar matrix. Transformed cells can then be either lysed and detected with a fluorescent dye or recovered and re-plated.

Easy Fluorescence Detection with the CytoSelect™ Cell Transformation Assay. HeLa and NIH3T3 cells were seeded at various concentrations and cultured for 6 days. Cell transformation was determined according to the assay protocol.

Cat. No.
DescriptionAmount
CBA-135
CytoSelect Cell Transformation Assay (Cell Recovery Compatible), Colorimetric
96 assays
CBA-135-5
CytoSelect Cell Transformation Assay (Cell Recovery Compatible), Colorimetric
5 x 96 assays
CBA-140
CytoSelect Cell Transformation Assay (Cell Recovery Compatible), Fluorometric
96 assays
CBA-140-5
CytoSelect Cell Transformation Assay (Cell Recovery Compatible), Fluorometric
5 x 96 assays
CBA-145
CytoSelect 384-well Cell Transformation Assay, Fluorometric
384 assays
CBA-145-5
CytoSelect 384-well Cell Transformation Assay, Fluorometric
5 x 384 assays

Recent Product Citations

  1. Hirata, H. et al. (2010). Role of secreted Frizzled-related protein3 in human renal cell carcinoma. Cancer Res. 70:1896-1905 (#CBA-135).
  2. Xie, G. et al. (2009). Acetylcholine-induced activation of M3 muscarinic receptors stimulates robust matrix metalloproteinase gene expression in human colon cancer cells. Am. J. Physiol. Gastrointest. Liver Physiol. 296:G755-G763 (#CBA-135).
  3. Hirata, H. et al. (2009). Wnt antagonist gene DKK2 is epigenetically silenced and inhibits renal cancer progression through apoptotic and cell cycle pathways. Clin. Cancer Res. 15:5678-5687 (#CBA-135).
  4. Ke, X-S. et al. (2008). Epithelial to mesenchymal transition of a primary prostate cell line with switches of cell adhesion modules but without malignant transformation. PLoS One 3(10):e3368 (#CBA-135).


Source:
Website: Cell Biolabs

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Brands: Cell Biolabs
Product groups: Assays