Fig.4. Effects of substrate on the ATPase activity of Hsc70 purified from Gillichthys mirabilis. On the x-axis, data sets 1–3 represent ATPase activity measurements for three independent Hsc70 preparations. Duplicate samples were incubated with 3.7x10⁵Bq of [-³²P]ATP at 23°C, and the amount of ATP hydrolysis was measured after 30min by thin-layer chromatography. Values are the mean pixel absorbance volume (background-corrected) + variance (N=2). Black, dark grey and light grey columns represent assays composed of 80µmoll^-1 RCMLA in assay buffer (AB), 2µg of Hsc70 in AB and 2µg of Hsc70 plus 80µmoll^-1 RCMLA in AB, respectively. See Materials and methods for a full description of the assay buffer components.