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Reporter Gene Assay

Beta-gal activity assay using DDAOG as substrate.

Figure 1. β-gal activity assay using DDAOG as substrate. HEK293 cells were transiently transfected with either a β-gal expressing vector (CMV-gal) or control vector. The signal intensity produced by DDAOG cleavage is linearly related to the substrate concentration. Signal intensities obtained using different concentrations of DDAOG (μM) were determined by imaging in the Odyssey 700 nm channel. The original image (A) and quantified data (B) are shown.

The β-galactosidase (β-gal) reporter gene is widely used in genetics and molecular biology. Uses include:

  • Monitoring promoter activity in transfected cells
  • Analyzing protein interactions and translocation using enzyme fragment complementation
  • Monitoring tumor growth in animal models1,2 (poor cellular penetration by the substrate affects performance in animals)

β-gal assays can be performed directly with the Odyssey® CLx, Odyssey Classic, or Odyssey Sa Infrared Imager in microplate format, using the near-infrared fluorescent substrate DDAOG (9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) β-D-galactopyranoside). The cleaved substrate fluoresces strongly in the 700 nm channel. This assay was published in Analytical Biochemistry in 2009.3

  • 1. Buller, C.J. et al. Methods Find Exp Clin Pharmacol. 25(9): 713-6
  • 2. Tung, C.H. et al. Cancer Res. 64(5): 1579-83 (2004)
  • 3. Gong, H. et al. Anal Biochem. 386(1): 59-64 (2009)

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