Hemolytic assay for human complement activity
Principle:
¡@¡@This expriment uses human complements in serum to lyse sheep erythrocytes, as an antibodies. If our herb's extract is useful, the lysis degree of erythrocytes will be minor.
Meterials & Methods:

¡@¡@Human pooled serum(HPS) was obtained from healthy rolunteers and stored at -20¢J until use.
¡@¡@Optimal serum concetration(control/100%lysis=1:40~1:60) was obtained by test before experiment.
¡@¡@Veronal saline buffer(VSB) containing 5mM veronal and 150mM NaCl at pH=7.4 was used. This served as a stock solution for VSB++ contain 0.5mM Mg2+ and 0.15mM Ca2+ and for EDTA-VSB containing 8mM EDTA.
¡@¡@Classical complement pathways activities were determined in HPS with uncoated rabbit antisheep erythrocytes antibodies and sensitized sheep erythrocytes as target antigens. All reagents were diluted in VSB++ and EDTA-VSB.
¡@¡@After washing sheep erythocytes(EDTA-VSB one time and VSB++ two times), herb's extract and serum(75£f:75£f) incubated on a shcker in 37¢J for 30 min. Then added 150£f sensitised sheep erythrocytes, and incubated in 37¢J for 30 min.
¡@¡@Finally centrifuge the solution at 1700rpm for 10 min, and drawed supernatant to detect it's absorbed value at 405nm.