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. 2002 Feb 26;99(5):2720–2725. doi: 10.1073/pnas.052436599

Figure 4.

Figure 4

The PS complex and γ-secretase activity can be eluted from the affinity resin. (a) Coimmunoprecipitates (Co-IP) were used to determine whether elution conditions disrupt PS heterodimers. CHAPSO-solubilized cell membranes were treated as indicated and then immunoprecipitated with X81 (to PS1 NTF). Precipitated PS1 NTF and PS1 CTF were detected by Western blotting. NP40, Nonidet P-40. (b) Solubilized γ-secretase was passed over a 31C affinity column, washed in binding buffer, and eluted in 1% Brij-35. The PS complex was coimmunoprecipitated with X81 and exchanged into 0.25% CHAPSO. This material was compared with the starting material in γ-secretase assays using N100Flag and C100Flag in the presence (+) or absence (−) of γ-secretase inhibitor 31C.