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Menene ayyukan kit ɗin ELISA?
2022-12-23
Kit ɗin ELISA ya dogara ne akan ingantaccen lokaci na antigen ko antibody da lakabin enzyme na antigen ko antibody. Antigen ko antibody da ke daure a saman madaidaicin mai ɗaukar hoto har yanzu yana riƙe da aikin sa na rigakafi, kuma enzyme da aka yiwa lakabin antigen ko antibody yana riƙe duka ayyukan rigakafi da aikin enzyme. A lokacin ƙaddamarwa, samfurin da aka gwada (wanda aka auna antibody ko antigen) yana amsawa tare da antigen ko antibody a saman babban mai ɗauka. Rukunin antigen-antibody da aka kafa akan ingantaccen mai ɗaukar hoto yana rabu da wasu abubuwa a cikin ruwa ta hanyar wankewa.
Ana ƙara antigens masu lakabin Enzyme ko ƙwayoyin rigakafi, waɗanda kuma suna ɗaure ga mai ƙarfi ta hanyar amsawa. A wannan lokacin, adadin enzyme a cikin lokaci mai ƙarfi yana daidai da adadin abu a cikin samfurin. Bayan ƙara da substrate na enzyme dauki, da substrate ne catalyzed da enzyme ya zama masu launi kayayyakin. Adadin samfurin yana da alaƙa kai tsaye da adadin abin da aka gwada a cikin samfurin, don haka ana iya aiwatar da ƙididdigar ƙima ko ƙididdiga bisa ga zurfin launi.
Babban haɓakar haɓakar ƙwayoyin enzymes a kaikaice yana haɓaka sakamakon amsawar rigakafi, yana mai da hankali sosai. Ana iya amfani da ELISA don ƙayyade antigens, amma kuma ana iya amfani da su don ƙayyade ƙwayoyin rigakafi.
Ka'idodin asali na kayan aikin ELISA
Yana amfani da takamaiman amsawar antigen da antibody don haɗa abu zuwa enzyme, sannan kuma yana samar da halayen launi tsakanin enzyme da substrate don tantance ƙididdigewa. Abinda ake aunawa zai iya zama antibody ko antigen.
Akwai reagents guda uku da ake buƙata a cikin wannan hanyar ƙaddara:
â Antigen mai ƙarfi ko antibody (adsorbent na rigakafi)
â¡ Enzyme mai lakabin antigen ko antibody (alama)
⢠don aikin enzyme (wakilin haɓaka launi)
A cikin ma'auni, antigen (antibody) an fara ɗaure shi da ƙarfi mai ɗaukar nauyi, amma har yanzu yana riƙe da aikin rigakafi, sa'an nan kuma an ƙara haɗin (alamar) na antibody (antigen) da enzyme, wanda har yanzu yana riƙe da ainihin aikinsa na rigakafi da enzyme. aiki. Lokacin da conjugate ya amsa tare da antigen (antibody) akan ingantaccen mai ɗaukar hoto, ana ƙara madaidaicin madaidaicin ƙwayar enzyme. Wato catalytic hydrolysis ko REDOX dauki da launi.
Ana ƙara antigens masu lakabin Enzyme ko ƙwayoyin rigakafi, waɗanda kuma suna ɗaure ga mai ƙarfi ta hanyar amsawa. A wannan lokacin, adadin enzyme a cikin lokaci mai ƙarfi yana daidai da adadin abu a cikin samfurin. Bayan ƙara da substrate na enzyme dauki, da substrate ne catalyzed da enzyme ya zama masu launi kayayyakin. Adadin samfurin yana da alaƙa kai tsaye da adadin abin da aka gwada a cikin samfurin, don haka ana iya aiwatar da ƙididdigar ƙima ko ƙididdiga bisa ga zurfin launi.
Babban haɓakar haɓakar ƙwayoyin enzymes a kaikaice yana haɓaka sakamakon amsawar rigakafi, yana mai da hankali sosai. Ana iya amfani da ELISA don ƙayyade antigens, amma kuma ana iya amfani da su don ƙayyade ƙwayoyin rigakafi.
Ka'idodin asali na kayan aikin ELISA
Yana amfani da takamaiman amsawar antigen da antibody don haɗa abu zuwa enzyme, sannan kuma yana samar da halayen launi tsakanin enzyme da substrate don tantance ƙididdigewa. Abinda ake aunawa zai iya zama antibody ko antigen.
Akwai reagents guda uku da ake buƙata a cikin wannan hanyar ƙaddara:
â Antigen mai ƙarfi ko antibody (adsorbent na rigakafi)
â¡ Enzyme mai lakabin antigen ko antibody (alama)
⢠don aikin enzyme (wakilin haɓaka launi)
A cikin ma'auni, antigen (antibody) an fara ɗaure shi da ƙarfi mai ɗaukar nauyi, amma har yanzu yana riƙe da aikin rigakafi, sa'an nan kuma an ƙara haɗin (alamar) na antibody (antigen) da enzyme, wanda har yanzu yana riƙe da ainihin aikinsa na rigakafi da enzyme. aiki. Lokacin da conjugate ya amsa tare da antigen (antibody) akan ingantaccen mai ɗaukar hoto, ana ƙara madaidaicin madaidaicin ƙwayar enzyme. Wato catalytic hydrolysis ko REDOX dauki da launi.
Inuwar launi da yake samarwa yana daidai da adadin antigen (antibody) da za a auna. Ana iya lura da wannan samfur mai launi ta ido tsirara, microscope na gani, microscope na lantarki, kuma ana iya auna shi ta spectrophotometer (kayan aikin lakabin enzyme). Hanyar yana da sauƙi, dacewa, sauri da takamaiman.
A baya:Gabatarwar Elisa Plate
