Fig. 2.

Tpn can form complexes with β₂m at physiological temperature. sTpn–A2 complexes were formed by incubating proteins (5 μM each) at 37°C either in the absence (A) or presence (B) of β₂m (50 μM) and immunoprecipitated with anti-FLAG. (C) sTpn (5 μM) and β₂m (50 μM), or β₂m alone were incubated for 2 h at 37°C, then immunoprecipitated with anti-FLAG. (A–C Upper) Silver-stained SDS/PAGE gels showing anti-FLAG IP of indicated proteins (A, lane 2; B, lanes 1 and 2; C lanes 1 and 2) or buffer (A, lane 1; C, lane 3) and direct protein loads of indicated concentration of A2 heterodimers (A and B, lanes 3–8). (A–C Lower) Anti-β₂m blot of the above gels. Data are representative of two independent analyses.