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Figure 1 | BMC Biochemistry

Figure 1

From: Limited proteolysis of human histone deacetylase 1

Figure 1

Limited proteolysis of endogenous and transiently expressed HDAC1. Immunoprecipitated endogenous HDAC1 (A) and transiently expressed HDAC1-Flag (HDAC1-F) (B) were incubated at room temperature with increasing concentrations of trypsin (0.000625 μg/μL- 1X, 0.00125 μg/μL- 2X, 0.0025 μg/μL- 4X, 0.005 μg/μL- 8X, 0.01 μg/μL- 16X, and 0.05 μg/μL- 80X). After separation by SDS-PAGE, the proteins were visualized with either anti-HDAC1 antibody in case of endogenous HDAC1 or anti-Flag antibody in case of transiently expressed HDAC1-F. (C) A graph showing percentage of full length endogenous HDAC1 (white) and transiently expressed HDAC1-Flag (black) remaining after exposure to different concentrations of trypsin. The curves were generated by least squares fit to a single exponential. (D) A plot displaying the deacetylase activity of HDAC1-F after incubation with an increasing concentration of trypsin. The inset displays the relationship between deacetylase activity and percentage of full length HDAC1-F remaining after exposure to increasing concentrations of trypsin and the data was fit to a linear equation.

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