TECHNICAL NOTE
8
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Analysis so ware (NanoTemper Technologies).
For the IMAC purifications the clarified lysates were applied to 50 μl Ni-NTA columns,
equilibrated with binding (50 mM Tris-HCl pH 8.0, 300 mM NaCl, 20 mM imidazole, 0.01%
(v/v) 1-thioglycerol). Next, the columns were washed with approx. 1 ml of binding buffer
and bound proteins eluted with 250 μl elution buffer (50 mM Tris-HCl pH 8.0, 300 mM
NaCl, 300 mM imidazole, 0.01% (v/v) 1-thioglycerol.
SDS-PAGE gels were runs according to Laemli2. Samples of the lysate, clarified lysate (10
μl + 30 μl loading buffer each) and the elution fraction (20 μl + 20 μl loading buffer) were
applied to the 15% acrylamide gels. The gels were stained with Page Blue protein staining
solution (Thermo Scientific) following the manufacturer's instructions.
