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. Author manuscript; available in PMC: 2015 Oct 21.
Published in final edited form as: Methods Enzymol. 2015 Apr 15;559:17–26. doi: 10.1016/bs.mie.2014.11.004

Step 1 Binding buffer.

Component Final concentration Stock Amount
Tris–HCl, pH 8.0 50 mM 1 M 50 ml
NaCl 200 mM 1 M 200 ml
EDTA 1 mM 0.5 M 2 ml

Add water to 1 l and pass through a 0.22 μm filter

Optional: 0.1-mM Na azide, or reducing agents such as 1 mM DTT or β-mercaptoethanol can be added