Figure 2
Isolation and immunodetection of the α 1 -AT /SP-A complex. A: Isolation of the complex by gel filtration chromatography on two Biosep SEC – S 4000 columns connected in series using HPLC. Gel filtration profiles: commercial α1-AT (in profile c; 19.32 ± 0.1 mL); purified SP-A (in profile b; 16.49 ± 0.07 mL); α1-AT /SP-A complex (in profile a; 15.31 ± 0.04 mL) and unreacted α1-AT (in profile a; 19.35 ± 0.09 mL). Inset: calibration curve obtained using the following standards: A = α2-macroglobulin (725 kDa), B = aldolase (158 kDa), C = bovine serum albumin (67 kDa), D = chymotrypsinogen (25 kDa), E = cytocrome C (12.5 kDa). B: Immunodetection of the complex. α1-AT was added to wells a1 and a2, peak 1 (α1-AT /SP-A complex) of Figure 2A was added to wells b1 and b2, and SP-A to wells c1 and c2. Antiserum anti-α1-AT was added to wells a1, b1 and c1, antiserum anti-SP-A was added to wells a2, b2 and c2. Peak 1 (α1-AT /SP-A complex) is recognized by both antisera.