Immunodetection of VHA-a in membrane and soluble fractions of M. crystallinum leaves. Membranes were isolated in the presence of 100 (1, 2), 500 mM NaCl (3, 4), and without (5) or with (6) protease inhibitor complete®, respectively. Membranes and soluble fraction were separated rapidly for (1)–(4) whereas the standard membrane isolation procedure with ultracentrifugation was undertaken for (5) and (6). The 95 kDa band is seen as the dominant protein in the membrane fraction isolated at high salt and to some part in the isolate obtained in the presence of protease inhibitor.
Kluge et al. BMC Cell Biology 2004 5:29 doi:10.1186/1471-2121-5-29