Soft Agar Colony Formation Assay: A Method to Test Effects of Novel Compounds on Cancer Cell Proliferation

Abstract

Source: Horibata A. et. al., Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells. J. Vis. Exp. (2015).

This video describes a soft agar colony formation assay to test the effects of a PADI enzyme inhibitor and BB-Cl-amidine on breast cancer tumorigenicity in vitro. This assay also enables the quantitative assessment of cell transformation potential within the context of genetic perturbations.

Protocol

1. Preparation of 3% 2-Hydroxyethyl Agarose Into a clean, dry 100 ml glass bottle, add 0.9 g of 2-hydroxyethyl agarose (Agarose VII) followed by 30 ml of distilled water. Microwave the mixture for 15 sec and gently swirl. Repeat this step at least three more times until the agarose powder fully dissolves. Autoclave the solution-containing bottle for 15 min. Allow the agarose solution to cool down to RT before further use. Store the solution at RT. <p class="j…

Materials

Zeiss Axiopot	Carl Zeiss Microscopy	1021859251
Inverted Microscope	Olympus	CKX41
DMEM/F-12	Lonza BioWhittaker	12-719F
HyClone Donor Equine Serum	Fisher Scientific	SH30074.03
Penicillin Streptomycin	Life Technologies	15140-122
2-Hydroxyethylagarose: Type VII, low gelling temperature	Sigma-Aldrich	39346-81-1