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Kodi ntchito za ELISA kit ndi ziti?
2022-12-23
ELISA zida zimatengera gawo lolimba la ma antigen kapena ma antibody ndi ma enzymes a antigen kapena antibody. Antigen kapena antibody yomangidwa pamwamba pa chonyamulira cholimba imasungabe mphamvu zake zoteteza chitetezo chathupi, ndipo enzyme yotchedwa antigen kapena antibody imasungabe ntchito yake ya immunological ndi ma enzyme. Panthawi yotsimikizira, chitsanzo chomwe chikuyesedwa (momwe antibody kapena antigen amayezedwa) chimakhudzidwa ndi antigen kapena antibody pamwamba pa chonyamulira cholimba. Antigen-antibody complex yomwe imapangidwa pa chonyamulira cholimba imasiyanitsidwa ndi zinthu zina zamadzimadzi mwa kuchapa.
Ma antigen olembedwa ndi ma enzyme kapena ma antibodies amawonjezeredwa, omwe amamangirizanso chonyamulira cholimba pochita. Panthawiyi, kuchuluka kwa enzyme mu gawo lolimba kumayenderana ndi kuchuluka kwa zinthu zomwe zili mu chitsanzocho. Pambuyo powonjezera gawo lapansi la momwe ma enzyme amachitira, gawo lapansili limapangidwa ndi enzyme kuti likhale zinthu zamitundumitundu. Kuchuluka kwa mankhwalawo kumagwirizana mwachindunji ndi kuchuluka kwa zinthu zomwe zayesedwa mu chitsanzo, kotero kuti kusanthula kwapamwamba kapena kuchuluka kungathe kuchitidwa molingana ndi kuya kwa mtundu.
Kuchita bwino kwambiri kwa ma enzymes mosadukiza kumakulitsa zotsatira za kuyankha kwa chitetezo chamthupi, kupangitsa kuyesako kukhala kosavuta. ELISA ingagwiritsidwe ntchito kudziwa ma antigen, komanso ingagwiritsidwe ntchito kudziwa ma antibodies.
Mfundo zoyambirira za ELISA kit
Imagwiritsa ntchito momwe ma antigen ndi ma antibody kuti alumikizitse chinthucho ndi enzyme, kenako amapanga mawonekedwe amtundu pakati pa enzyme ndi gawo lapansi kuti adziwe kuchuluka kwake. Chinthu choyezera chikhoza kukhala antibody kapena antigen.
Pali ma reagents atatu ofunikira pakutsimikiza uku:
â Maantijeni a gawo lolimba kapena ma antibody (immune adsorbent)
â¡ Enzyme yolembedwa kuti antigen kapena antibody (chikho)
⢠gawo lapansi la ma enzyme (wothandizira kukula kwamtundu)
Pakuyezera, antigen (antibody) imamangiriridwa koyamba ndi chonyamulira cholimba, koma imasungabe chitetezo chake, ndiyeno conjugate (chizindikiro) cha antibody (antigen) ndi enzyme imawonjezeredwa, yomwe imasungabe chitetezo chake choyambirira ndi enzyme. ntchito. Pamene conjugate imachita ndi antigen (antibody) pa chonyamulira cholimba, gawo lapansi lolingana la enzyme limawonjezeredwa. Ndiko kuti, catalytic hydrolysis kapena REDOX reaction ndi mtundu.
Ma antigen olembedwa ndi ma enzyme kapena ma antibodies amawonjezeredwa, omwe amamangirizanso chonyamulira cholimba pochita. Panthawiyi, kuchuluka kwa enzyme mu gawo lolimba kumayenderana ndi kuchuluka kwa zinthu zomwe zili mu chitsanzocho. Pambuyo powonjezera gawo lapansi la momwe ma enzyme amachitira, gawo lapansili limapangidwa ndi enzyme kuti likhale zinthu zamitundumitundu. Kuchuluka kwa mankhwalawo kumagwirizana mwachindunji ndi kuchuluka kwa zinthu zomwe zayesedwa mu chitsanzo, kotero kuti kusanthula kwapamwamba kapena kuchuluka kungathe kuchitidwa molingana ndi kuya kwa mtundu.
Kuchita bwino kwambiri kwa ma enzymes mosadukiza kumakulitsa zotsatira za kuyankha kwa chitetezo chamthupi, kupangitsa kuyesako kukhala kosavuta. ELISA ingagwiritsidwe ntchito kudziwa ma antigen, komanso ingagwiritsidwe ntchito kudziwa ma antibodies.
Mfundo zoyambirira za ELISA kit
Imagwiritsa ntchito momwe ma antigen ndi ma antibody kuti alumikizitse chinthucho ndi enzyme, kenako amapanga mawonekedwe amtundu pakati pa enzyme ndi gawo lapansi kuti adziwe kuchuluka kwake. Chinthu choyezera chikhoza kukhala antibody kapena antigen.
Pali ma reagents atatu ofunikira pakutsimikiza uku:
â Maantijeni a gawo lolimba kapena ma antibody (immune adsorbent)
â¡ Enzyme yolembedwa kuti antigen kapena antibody (chikho)
⢠gawo lapansi la ma enzyme (wothandizira kukula kwamtundu)
Pakuyezera, antigen (antibody) imamangiriridwa koyamba ndi chonyamulira cholimba, koma imasungabe chitetezo chake, ndiyeno conjugate (chizindikiro) cha antibody (antigen) ndi enzyme imawonjezeredwa, yomwe imasungabe chitetezo chake choyambirira ndi enzyme. ntchito. Pamene conjugate imachita ndi antigen (antibody) pa chonyamulira cholimba, gawo lapansi lolingana la enzyme limawonjezeredwa. Ndiko kuti, catalytic hydrolysis kapena REDOX reaction ndi mtundu.
Mthunzi wamtundu womwe umatulutsa umagwirizana ndi kuchuluka kwa ma antigen (antibody) oti ayezedwe. Chojambula chachikuda ichi chikhoza kuwonedwa ndi maso, maikulosikopu ya kuwala, maikulosikopu ya elekitironi, imathanso kuyesedwa ndi spectrophotometer (chida cholembera enzyme). Njirayi ndi yosavuta, yabwino, yofulumira komanso yeniyeni.
Zam'mbuyo:Cotaus akukuyembekezerani ku 2023EBC!
