. Author manuscript; available in PMC: 2020 May 13.

Published in final edited form as: J Vis Exp. 2020 Mar 29;(157):10.3791/61018. doi: 10.3791/61018

Table 1: Solutions used for APEGS assay.

Homogenization buffer and buffer A can be prepared ahead of time; however, add protease inhibitors and phenylmethylsulfonyl fluoride (PMSF) immediately before use. All other solutions should be prepared fresh.

Buffer	Components	Working conc.	Comments
Homogenization	0.32 M sucrose, 1 mM Tris pH 7.2, 1 mM MgCl₂		Add PMSF and protease inhibitors immediately before use.
Buffer A	PBS pH 7.2, 5 mM EDTA, 4% SDS (w/v)		Add PMSF and protease inhibitors immediately before use.
TCEP solution	500 mM TCEP in ddH₂O (pH 7.2)	25 mM	Add 10 N NaOH to increase pH.
NEM solution	2.0 M NEM in 100% ethanol	50 mM	Prepare fresh.
Buffer H (HAM+)	1.33 M HAM, pH 7.0, 5 mM EDTA, 0.2% Triton X-100 (w/v)	1.0 M	Prepare fresh.
Buffer T (HAM−)	1.33 M Tris-HCl, pH 7.0, 5 mM EDTA, 0.2% Triton X-100 (w/v)
mPEG-5k	100 mM mPEG-5k in ddH₂O	20 mM	Mix well. Highly viscous.