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Fig. 3 | BMC Biochemistry

Fig. 3

From: Identification and characterization of the novel nuclease activity of human phospholipid scramblase 1

Fig. 3

Nuclease properties of hPLSCR1. a Nuclease assay for hPLSCR1 with yeast and human genomic DNA (gel assay): Nuclease assay was performed as described in ‘Methods’ section. 20 pmol of hPLSCR1 (enzyme) was treated with 200 ng of yeast and human genomic DNA (substrate) in assay buffer at 37 °C for 60 min and visualized on 1 % agarose gel with ethidium bromide. b Native PAGE for visualization of nuclease activity (gel assay): 200 ng of human genomic DNA was incubated with hPLSCR1 at 37 °C for 60 min and visualized on a 12 % native PAGE stained with 0.5 μg/ml ethidium bromide. ‘Ladder’ denotes 1 kb ladder. c Time dependent nuclease activity of hPLSCR1 (gel assay): hPLSCR1 (20 pmol) was incubated with human genomic DNA (200 ng) at 37 °C for 5, 10, 15, 30, 60, 90 min along with a no-enzyme control (control) and visualized on 1 % agarose gel with ethidium bromide. d Effect of substrates on nuclease activity (gel assay): Nuclease assays were performed with 200 ng of plasmid DNA and total RNA as substrates along with a no-enzyme control visualized on a 1 % agarose gel. e Nuclease activity of hPLSCR1 with ssDNA (gel assay): Nuclease assays were performed with 200 ng of a random ssDNA (43 bases) and dsDNA (human genomic DNA) along with a no-enzyme control containing both the substrates and visualized on a 2 % agarose gel

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