溶血素的收集和溶血素單位的滴定[1]

1、溶血素的收集和溶血素單位的滴定1實(shí)驗(yàn)四實(shí)驗(yàn)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1補(bǔ)體結(jié)合位點(diǎn)補(bǔ)體結(jié)合位點(diǎn)溶血素的收集和溶血素單位的滴定1Fab段段Fc段段C1q 結(jié)合結(jié)合位點(diǎn)被屏位點(diǎn)被屏障障結(jié)合抗原之前結(jié)合抗原之前 CH1 CH2暴露的暴露的C1q結(jié)結(jié)合位點(diǎn)合位點(diǎn)結(jié)合抗原之后結(jié)合抗原之后IgM CH3區(qū)，區(qū)，IgG CH2區(qū)區(qū) 原理原理溶血素的收集和溶血素單位的滴定1Ag + AbAg + AbComplementAg + Ab + Ag + Ab + C C補(bǔ)體活化補(bǔ)體活化 原理原理溶血素的收集和溶血素單位的滴定1補(bǔ)體活化補(bǔ)體活化裂解

2、裂解 原理原理溶血素的收集和溶血素單位的滴定1SRBCSRBCMouseMouseAnti-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1SRBCSRBC免疫小鼠免疫小鼠2%SRBC 2%SRBC 補(bǔ)體補(bǔ)體(1:30) (1:30) 材料材料Test 1 Determination of the titer of hemolysinTest 1 Determination of the titer o

3、f hemolysin溶血素的收集和溶血素單位的滴定1 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?qū)⒀菏占翆⒀菏占罞pEp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.

4、1ml0.1ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml0.25ml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1123456N.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

5、.2ml(Original)(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(jié)果觀察結(jié)果7- - - - - - - 0.2ml0.

6、2ml0.6ml0.6ml0.2ml0.2ml溶血素的收集和溶血素單位的滴定1 結(jié)果結(jié)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(wú)溶血無(wú)溶血( (對(duì)照對(duì)照) )溶血素的收集和溶血素單位的滴定1 題目題目: 經(jīng)乙肝病毒表面抗原（HBsAg ）免疫的個(gè)體，如何定量檢測(cè)相應(yīng)特異性抗體的產(chǎn)生情況？ 目的，方法，預(yù)期結(jié)果，討論 溶血素的收集和溶血素單位的滴定108&5lgG3j)WyL2k698OEt&SIda

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