Clonogenic Assays

Creative Bioarray has experienced scientific team and state-of-the-art platform to provide reliable clonogenic assays for our customers worldwide. We are your ideal partner in fundamental research and drug development.

Introduction of Clonogenic Assays

Clonogenic assay, also known as colony formation assay, is an in vitro cell survival assay, which evaluates all modalities of cell death according to the ability of a single cell to grow into a colony. The assay essentially determines every cell in the population for its ability to proliferate indefinitely. The experiment mainly involves four major steps, including cell culture preparation, cell plating and treatment, fixation and staining, and data analysis from colony counts.

Clonogenic assay, which was originally developed to study the effect of radiation on cells, have played an essential role in radiobiology. At present, they have been widely used to determine the effectiveness of cytotoxic agents or various treatments with potential clinical application value, as well as in the field of cancer research to assess the survival and proliferative capability of cells, tumor-initiating capacity and the status of stem cells. Compared with cell viability assays and apoptosis assays, clonogenic assays can measure long-term effects of anti-cancer treatments.

Clonogenic assay performed in six-well plates, with clones produced by SW-1573 lung tumor cells. Figure 1. Clonogenic assay performed in six-well plates, with clones produced by SW-1573 lung tumor cells. (Franken N A P, et al., 2006)

Clonogenic Assays Available at Creative Bioarray

Basic Clonogenic Assay

Basic clonogenic assay is suitable for providing information regarding anchorage-dependent growth. Adherent cells are seeded on a cell culture plate to measure the ability of cells to form colonies. The key to success in this assay is the preperation of cell suspension and cell inoculation density.

Automated Clonogenic Assay

Basic clonogenic assay can be automated by using image cytometry and integrated plate stacker, thus saving a lot of time and cost. Image analyzer can be used for standardized analysis of the measured colony size and number. Automated clonogenic assay can be applied to different cell culture plates, from 6 well-plates to 96-well microplates.

Soft Agar Colony Formation Assay

Soft agar colony formation assay, developed on the basis of basic clonogenic assay, is a well-established method to assess the anchorage-independent growth ability of cells. It is widely used in the evaluation of cell transformation and tumorigenicity in vitro. Soft agar colony formation assay can also be used in the efficacy evaluation of anti-tumor therapeutics. Fluorometric dye and specialized agar solution can be used to modify this assay.

Creative Bioarray can provide customized services for individual requirements. We have built a reputation by providing high-quality results. If you need more detailed information, please feel free to contact us. We look forward to cooperating with you.


  1. Franken N A P, et al. Clonogenic assay of cells in vitro. Nature protocols, 2006, 1(5): pp: 2315-2319.
  2. Rafehi H, et al. Clonogenic assay: adherent cells. JoVE (Journal of Visualized Experiments), 2011, 49: e2573.
  3. Nuryadi E, et al. Inter-assay precision of clonogenic assays for radiosensitivity in cancer cell line A549. Oncotarget, 2018, 9(17), pp: 13706-13712.
  4. Horibata S, et al.  Utilization of the soft agar colony formation assay to identify inhibitors of tumorigenicity in breast cancer cells. JoVE (Journal of Visualized Experiments), 2015, 99: e52727.
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