1、溶血素的收集和溶血素單位的滴定實驗四實驗四 溶血素的收集和溶血素溶血素的收集和溶血素單位的滴定單位的滴定溶血素的收集和溶血素單位的滴定補體結合位點補體結合位點溶血素的收集和溶血素單位的滴定Fab段段Fc段段C1q 結合結合位點被屏位點被屏障障結合抗原之前結合抗原之前 CH1 CH2暴露的暴露的C1q結結合位點合位點結合抗原之后結合抗原之后IgM CH3區，區，IgG CH2區區 原理原理溶血素的收集和溶血素單位的滴定Ag + AbAg + AbComplementAg + Ab + Ag + Ab + C C補體活化補體活化 原理原理溶血素的收集和溶血素單位的滴定補體活化補體活化裂解裂解 原理

2、原理溶血素的收集和溶血素單位的滴定SRBCSRBCMouseMouseAnti-SRBC AbAnti-SRBC Ab(hemolysin)(hemolysin) 免疫免疫3 times / 3ws3 times / 3wsIgGIgGHbHbHbHbHbHbSerumSerum 原理原理溶血素的收集和溶血素單位的滴定SRBCSRBC免疫小鼠免疫小鼠2%SRBC 2%SRBC 補體補體(1:30) (1:30) 材料材料Test 1 Determination of the titer of hemolysinTest 1 Determination of the titer of hemol

3、ysin溶血素的收集和溶血素單位的滴定 方法方法1. 1. 收集血清收集血清摘取摘取SRBCSRBC免疫小鼠眼球免疫小鼠眼球將血液收集至將血液收集至EpEp管中管中4 4 放置放置30min30min12000rpm 12000rpm 離心離心10min10min取上層血清取上層血清1 12 23 34 45 5系列稀釋系列稀釋1:5,1:5, 1:10, 1:20, 1:40, 1:801:10, 1:20, 1:40, 1:80血清血清N.SN.S0.4ml0.4ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml原液原液0.1ml0.1ml

4、0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml稀釋比例稀釋比例1:51:51:101:101:201:201:401:401:801:80mixmixmixmixmixmixmixmix溶血素的收集和溶血素單位的滴定123456N.SN.S系列稀釋系列稀釋血清血清ComplementComplement0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml徹底混勻徹底混勻3737, 30min, 30min2. 2. 溶血素單位的滴定溶血素單位的滴定0.2ml0.2ml(Orig

5、inal)(Original)0.2ml0.2ml(1:5)(1:5)0.2ml0.2ml(1:10)(1:10)0.2ml0.2ml(1:20)(1:20)0.2ml0.2ml(1:40)(1:40)0.2ml0.2ml(1:80)(1:80)2%SRBC2%SRBC0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.2ml0.4ml0.4ml0.4ml0.4ml0.4ml0.4ml0.4ml0.4ml0.4ml0.4ml0.4ml0.4ml3. 3. 觀察結果觀察結果7- - - - - - - 0.2ml0.2ml0.6ml0.6ml0.2ml0.2ml溶血素的收集和溶血素單位的滴定 結果結果Test 1 Determination of the titer of hemolysinTest 1 Determination of the titer of hemolysin完全溶血完全溶血不完全溶血不完全溶血無溶血無溶血(