schistosoma elisa酶(mei)聯免疫試(shi)劑盒樣本實(shi)驗前(qian)準(zhun)備(bei):
ELISA試(shi)劑盒液(ye)體(ti)樣本:包括血(xue)清、血(xue)漿(jiang)、尿(niao)液(ye)、胸(xiong)腹(fu)水(shui)、腦脊(ji)液(ye)、細(xi)胞培(pei)養上(shang)清等(deng)。
()血(xue)清
室溫(wen)血(xue)液(ye)自然凝固(gu)0-20分(fen)鐘(zhong)後(hou),離心20分(fen)鐘(zhong)左(zuo)右(2000-3000轉(zhuan)/分(fen))。仔(zai)細(xi)收(shou)集上清。保存過(guo)程中(zhong)如(ru)有沈(chen)澱形(xing)成,應再(zai)次(ci)離心。
(2)血(xue)漿(jiang):
應根據標(biao)本(ben)的(de)要(yao)求選(xuan)擇EDTA、檸檬酸(suan)鈉或肝(gan)素作為抗凝(ning)劑,混合0-20分(fen)鐘(zhong)後(hou),離心20分(fen)鐘(zhong)左(zuo)右(2000-3000轉(zhuan)/分(fen))。仔(zai)細(xi)收(shou)集上清。保存過(guo)程中(zhong)如(ru)有沈(chen)澱形(xing)成,應再(zai)次(ci)離心。
(3)尿(niao)液(ye):
用無(wu)菌(jun)管(guan)收(shou)集(ji)。離心20分(fen)鐘(zhong)左(zuo)右(2000-3000轉(zhuan)/分(fen))。仔(zai)細(xi)收(shou)集上清。保存過(guo)程中(zhong)如(ru)有沈(chen)澱形(xing)成,應再(zai)次(ci)離心。胸腹(fu)水(shui)、腦脊(ji)液(ye)參(can)照此實(shi)行。
(4)細(xi)胞培(pei)養上(shang)清:
檢測分(fen)泌(mi)性(xing)的(de)成(cheng)份(fen)時(shi),用無(wu)菌(jun)管(guan)收(shou)集(ji)。離心20分(fen)鐘(zhong)左(zuo)右(2000-3000轉(zhuan)/分(fen))。仔(zai)細(xi)收(shou)集上清。
(5)培(pei)養細(xi)胞
檢測細(xi)胞內的成份(fen)時(shi),用PBS(PH7.2-7.4)稀(xi)釋(shi)細(xi)胞懸(xuan)液(ye),細(xi)胞濃(nong)度(du)達到00萬(wan)/ml左右(you)。通過(guo)反(fan)復(fu)凍(dong)融(rong)或加(jia)入組(zu)織蛋白(bai)萃取(qu)試(shi)劑,以使(shi)細(xi)胞破(po)壞並(bing)放(fang)出(chu)細(xi)胞內成份。離心20分(fen)鐘(zhong)左(zuo)右(2000-3000轉(zhuan)/分(fen))。仔(zai)細(xi)收(shou)集上清。保存過(guo)程中(zhong)如(ru)有沈(chen)澱形(xing)成,應再(zai)次(ci)離心。
(6)組織標(biao)本(ben)
切割(ge)標(biao)本(ben)後(hou),稱(cheng)取(qu)重(zhong)量。加(jia)入壹(yi)定(ding)量的(de)PBS,PH7.4。用液(ye)氮(dan)迅(xun)速冷(leng)凍(dong)保存備(bei)用。標(biao)本(ben)融(rong)化後(hou)仍(reng)然保持2-8℃的(de)溫(wen)度(du)。加(jia)入壹(yi)定(ding)量的(de)PBS(PH7.4),或組織蛋白(bai)萃取(qu)試(shi)劑, 用手(shou)工或勻漿(jiang)器將(jiang)標(biao)本(ben)勻漿(jiang)化。離心20分(fen)鐘(zhong)左(zuo)右(2000-3000轉(zhuan)/分(fen))。仔(zai)細(xi)收(shou)集上清。分(fen)裝(zhuang)後(hou)壹(yi)份待檢測,其(qi)余(yu)冷(leng)凍(dong)備(bei)用。
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