Hielscher faka'ata tekinolosia

ʻOku pehe ʻe ultrasonic sipinga naʻe teuteuʻi maʻa ʻELISA As

ʻOku pehe ʻe ʻELISA ʻoku lahi hono fakaʻaongaʻi ki he diagnostics, fakatokanga ki he mahaki mo e mapuleʻi lelei (e.g. vakaiʻi ʻo e meʻakai allergens). Ko e sipinga ʻo e teuteu ultrasonic ko ha founga vave, falalaʻanga mo reproducible ia ke lyse toʻotoʻo mo fakamavaheʻi intracellular polotini, DNA, RNA mo organelles. ʻOku ʻomi ʻe Hielscher Ultrasonics ha ngaahi founga ultrasonic kehekehe ki hono teuteuʻi lelei ʻo e ngaahi sipinga tautaha, ngaahi meʻa kehekehe pea pehe ki microtiter ʻu lauʻi peleti mo e ʻu lauʻi peleti lelei 96.

ʻILISA – Enzyme-fehokotaki Immunosorbent as

ʻOku taukaveʻi ʻe ʻELISA ʻa e lea enzyme ʻoku fehokotaki mo e immunosorbent pea ko ha founga ia ʻoku fakaʻaongaʻi lahi biochemical ki he faʻahinga ʻo e ligand ʻoku haʻi ʻaki. ʻI he ELISA, ʻoku tanaki atu ha sipinga huhuʻa ki ha konga ʻoku tuʻu maʻu mo ha ngaahi koloa makehe. ʻI he angamaheni, ʻoku fakaʻaongaʻi ʻa e konga fefeka ko ʻeni ko ha coating ki ha peleti lelei pe ko e peleti ʻa ʻELISA. ʻI he taimi ko ia, ʻoku tanaki atu leva ha reagents huhuʻa kehekehe, incubated, mo fufulu, koeʻuhi ke faifai pea hoko ha liliu (e.g., fakalakalaka ʻo e lanu ʻaki e ola ʻo ha tali enzymatic) ʻoku hoko ia ʻi he vai fakaʻosi ʻi he vaitupu. ʻOku fakaʻata ʻe he liliu ko ʻeni ke fuatautau ʻa e lahi ʻo e analyte ʻaki ha quantitative “laukonga ". ʻOku fakaʻaongaʻi ʻa e quantitative laukonga, spectrophotometer, fluorometer, pe luminometer ke ʻiloʻi mo fuatautau ʻaki ʻa e malohi ʻo e maama ʻoku ʻave. ʻOku takiekina ʻa e ongoʻingofua ʻo e fakatokanga ʻe he amplification ʻo e fakaʻilonga lolotonga e ngaahi toʻonga analytic. Koeʻuhi ʻoku fakatotoloʻi lelei mo falalaʻanga ʻa e ngaahi toʻonga enzyme amplification, ʻoku fakaʻaongaʻi leva ʻa e enzymes ke faʻu ʻaki ʻa e fakaʻilonga. ʻOku fehokotaki ʻa e enzymes mo e fakatokanga reagents ʻi ha ngaahi kongokonga pau ke fakaʻata ʻa e fakamatala ʻoku tonu, ʻa ia ʻoku ne fakamatalaʻi foki ʻa e hingoa ko e "enzyme-fehokotaki" immunosorbent lea.
Hange ko ia ʻoku pehe ʻe ʻELISA ʻoku fakahoko ʻi microtiter ʻu lauʻi peleti/96-ʻu lauʻi peleti lelei, ʻoku ʻiloa ia ko e founga ʻoku fakatefito ʻi he peleti pea ʻoku fakaʻaongaʻi ia ʻi he kiliniki diagnostic, fakatotolo, fakalakalaka ʻo e faitoʻo konatapu mo e ala meʻa pehe. ki hono fakaʻilo mo quantifying maluEi, peptides, polotini, mo e homouni.
ʻOku faʻa fakaʻaongaʻi ʻa e founga ʻa ʻELISA ko ha meʻangaue diagnostic ʻi he faitoʻo, kautaha biotech ʻi, to pathology pea ko ha meʻafua lelei foki ia ʻi he ngaahi ngaueʻanga kehekehe.

Fakakakato 'a e fokotu'utu'u 'o e VialTweeter: VialTweeter sonotrode 'i he ultrasonic processor UP200St

Naʻe teuteuʻi ʻe he sipinga ultrasonic e ʻiuniti VialTweeter ʻOku fakaʻaongaʻi ia ki he lysis toʻotoʻo mo e ivi toʻo hingoa kimuʻa pea pehe ʻe ʻELISA

Kole 'a e fakamatala




Fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


Ultrasonic sipinga teuteuʻi kimuʻa ʻi he ʻELISA

Kimuʻa pea lava ke fakahoko ʻa e lea ʻa ʻELISA, ʻoku fie maʻu ʻe he sipinga ha ngaahi sitepu ki hono teuteuʻi ʻo e lysis toʻotoʻo mo e toʻo ʻuli ʻo e intracellular polotini, DNA, RNA mo e ala meʻa pehe. Ko e faingamalie ʻo ultrasonic lysis toʻotoʻo mo e ivi mavahe ko ʻene lavameʻa, falalaʻanga mo e reproducibility. ʻOku mahuʻinga ʻa e ngaahi moʻoniʻi meʻa kotoa ko ʻeni kae lava ke maʻu ʻa e diagnostics lelei mo e ngaahi ola ʻo e fekumi

Ngaahi lelei ʻo ultrasonic Lysis ʻi he ʻao ʻo ʻELISA

  • Homogeneous sipinga ʻo e faitoʻo
  • Fakakakato ʻa e lysis
  • Fakakakato e ivi ke toʻo ʻaki e ʻulí (hangē ko e maluEi, DNA)
  • Tokolahi lelei fulihi ki he faʻahinga toʻotoʻo
  • Ki ha faʻahinga sipinga pe ʻo e size
  • ngaahi
  • Mapuleʻi ʻo e ʻea
  • Fakamatala fakahangatonu protocoling ʻi he SD-card

Protocol ki he teuteu ʻa ʻELISA ultrasonic toʻotoʻo Lysis

Vakili-fa'ahinga insonifier UP200St ki he lysis

  • Ki he ngaahi anga fakafonua toʻotoʻo: Kimuʻa ʻi he ultrasonic toʻotoʻo lysis, centrifuge e ngaahi selo ʻi ha miniti ʻe 5 ʻi he 270 x g ʻi ha microcentrifuge. Toʻo ʻa e supernatant ʻaki ʻa e fakaʻamu mo resuspend ngaahi selo ʻi he 30 – 100 μL ʻo e RIPA ngataʻanga. ʻI he taimi ko ia, incubate ʻa e pellet toʻotoʻo ʻi he ʻaisi ʻi ha miniti ʻe 30.
  • ʻI he taimi ni, ʻoku mateuteu ʻa e sipinga toʻotoʻo ki ultrasonic lysis:
    Fakaʻaongaʻi ha fekumi-type ultrasonicator (e.g. UP200Ht mo e S26d2 fekumi) pe ko ha meʻangaue ultrasonic fakavahaʻa-taʻu (e.g. VialTweeter ki hoko sonication ʻo aʻu ki he ngaahi peleti ʻe 10 pe ko e UIP400MPT ki microtiter ʻu lauʻi peleti/96-ʻu lauʻi peleti) ʻo fakatatau mo e lahi ʻo e ngaahi sipinga ʻoku fie maʻu ke ke teuteu.
    Ke maʻu ʻa e fekumi-taipeʻi sonication ʻo ha sipinga pe ʻe taha, fokotuʻu ʻa e ngaahi selo ʻi he 1.5 mL microcentrifuge tiupi.
  • Tomuʻa fokotuʻu hoʻo ultrasonic taimi, fakahu fakakatoa ʻo e ivi, siakale ʻo e mode mo e/pe mafana ʻo e ʻea ʻi he menu fakaʻilekitulonika ʻo e ultrasonicator. ʻOku fakapapauʻi ʻe he meʻa ni ʻoku falalaʻanga ʻaupito sonication mo repeatability.
  • Fokotuʻu ʻa e sonotrode pea fetongi ʻa e ultrasonic device ʻi he. ʻUnuakiʻi fakalelei ʻa e kiʻi tokoni Micro ʻo e ultrasonic fekumi ʻo fakafou ʻi he sipinga ke sonicate ʻa e sipinga uniformly.
    Ko e lahi taha ʻo e ngaahi selo, ʻe fakakakato ultrasonic lysis hili ha siakale ʻe 2-4 ʻo e 10 Sec sonication.
  • Hili sonication, toʻo ʻa e sonotrode mei he sipinga. ʻOku totonu ke incubated ʻa e ngaahi sipinga ʻi he ʻaisi ʻi ha miniti ʻe 5. ʻI he taimi ko ia, centrifuge ʻi he 10,000 x g ʻi ha miniti ʻe 20 ke pellet ʻa e veve. ʻAve ʻa e supernatants ki ha foʻi tiupi microcentrifuge foʻou. Fakahingoa ʻa e analytes mo e falekoloa ʻi he-20 °C.
  • ʻE lava ke fakamaʻa ʻa e ultrasonic sonotrode ʻaki hano holoholoʻi fakalelei ia ʻaki ʻa e kava malohi pe sonicate ʻi ha beaker fonu ʻi he kava malohi, e.g. 70% ethanol. ʻOku autoclavable ʻa e ultrasonic probes kotoa pe naʻe ngaohi mei titanium.

Ki he homogenates moluu:

  • Fufulu ʻa e molu ʻaki ʻa e PBS momoko (0.01 M, pH = 7.4) ke toʻo fakaʻauliliki ʻa e toto hemolysis.
  • Fakafuofuaʻi ʻa e molu (kofuua, loto, maʻamaʻa, spleen mo e ala meʻa pehe) pea macerate ia ki ha fanga kiʻi kongokonga iiki, ʻa ia ʻoku homogenized ʻi PBS. ʻOku fekauʻaki ʻa e voliume ʻo e PBS ʻoku fie maʻu mo e mamafa ʻo e molu. ʻI heʻene hoko ko e pule ʻo e motuʻa tuhu, 1g ʻoku fie maʻu ki ai ha approx. 9mL PBS. ʻOku fokotuʻu atu ke tanaki atu ha protease inhibitor ki he PBS. (RIPA pe hypotonic lysis ngataʻanga ʻoku ʻi ai e protease mo e phosphatase inhibitor ʻe lava ke fakaʻaongaʻi pe.)
  • Fakatatau mo e lahi ʻo e vortex, ʻe lava ke tokoni ha faitoʻo nounou (approx miniti ʻe 1-2. ʻi he 15 Sec. pulses) ke teuteuʻi ʻa e foʻi molu.
  • Fakapipiki ha kiʻi tokoni Micro, e.g. S26d2, ki hoʻo ultrasonicator. Tuku ʻa e sipinga ʻo e foʻi leta ʻaki ʻa e foʻi leta ʻi ha falekaukau ʻaisi.
  • Sonicate ʻa e sipinga ʻaki hoʻo ultrasonicator, e.g. UP200St (80% amplitude) ʻi ha miniti ʻe 1. ʻi he founga ta hono mafu (15sec on, 15sec kiʻi malolo). Tauhi ʻa e sipinga ʻi he ʻaisi kaukau.
  • ʻOku centrifuged leva ʻa e homogenates ke maʻu ha ngaahi luo pau (cytosolic, ivi fakaʻatomi, mitochondrial pe lysosomal) kae lava ke fakalahi ʻa e ivi ki hono ʻanalaiso. ʻI centrifuging ʻa e sipinga ʻi ha miniti ʻe 5 ʻi he 5000×g, ʻoku toe ʻomi ai ʻa e supernatant.
ʻOku ʻi ai ha ultrasonic ʻe valu ʻi he sonotrode MTP-24-8-96 probes ki he sonication ʻo e ngaahi peleti microtiter.

ʻOku ʻi ai ha ultrasonic ʻe valu ʻi he sonotrode MTP-24-8-96 probes ki he sonication ʻo e ngaahi peleti microtiter.

Mapuleʻi falalaʻanga ʻo e ʻea lolotonga Sonication

Ko e mafana ʻo e ʻea ko ha meʻa mahuʻinga ia ʻoku matuʻaki mahuʻinga ki hono faitoʻo ʻo e ngaahi sipinga totonu, hange ko e ke taʻofi hono fakafuofuaʻi ʻo e polotini. ʻI he hoko ʻa e sipinga fakamisini kotoa pe, ʻoku fakatupu ʻe sonication ha vela. Neongo ia, ʻe lava ke mapuleʻi lelei e mafana ʻo e ngaahi sipinga ʻi he taimi ʻoku fakaʻaongaʻi ai e ngaahi device Hielscher Ultrasonics. ʻOku mau ʻoatu ha ngaahi fili kehekehe ke mou vakaiʻi mo puleʻi ʻaki e mafana hoʻomou ngaahi sipinga lolotonga hono teuteuʻi kinautolu ʻaki ha fekumi-type ultrasonicator pe ko e VialTweeter kimuʻa he analytically.

  1. Vakaiʻi ʻa e sipinga ʻo e ʻea: ʻOku fakanaunau ʻa e Hielscher digital ultrasonic processors ʻaki ha polokalama fakakomipiuta fakapotopoto mo ha mafana pluggable sensor resistance. Palaki ʻa e mafana ʻo e ʻea sensor resistance ki he meʻangaue ultrasonic (e.g., UP200Ht, UP200St, VialTweeter, UIP400MTP) pea fakahu ʻa e tokoni ʻa e ʻea sensor resistance ʻi he taha ʻo e ngaahi sipinga ʻo e tiupi. Te ke lava ʻo fokotuʻu ʻi he menu ʻo e ultrasonic processor ha mafana pau ki hoʻo sipinga sonication, ʻo fakafou ʻi he digital coloured Touch. ʻE taʻofi leva ʻa e ultrasonicator ʻi he taimi ʻe aʻu ai ʻa e mafana ʻo e ʻea pea kiʻi malolo kae ʻoua kuo aʻu ʻa e sipinga ʻo e ʻea ki he tuʻunga ʻulungaanga mahuʻinga maʻulalo ange ʻo e mafana ʻo e ʻ∆ mafana. Pea toe kamata leva ʻa e sonication. ʻOku taʻofi ʻe he founga fakapotopoto ko ʻeni ʻa e maʻulalo ʻoku ne fakaʻaiʻai ʻa e ʻea.
  2. Fekauʻaki mo e ultrasonic fakavahaʻa-taʻu ʻa e ʻiuniti VialTweeter, ko e konga titanium, ʻa ia ʻoku ne maʻu ʻa e sipinga ʻo e tiupi, ʻe lava ke tomuʻa ʻita. Faʻo ʻa e konga VialTweeter (ko e sonotrode pe taʻe transducer!) ki he ʻaisi pe ʻaisi ke teuteu ke mokomoko ʻa e konga titanium ʻoku tokoni ke toloi ʻa e mafana ʻo e ʻea ʻi he sipinga. Kapau ʻe lava, ʻe lava ke tomuʻa ʻita foki mo e sipinga.
  3. Fakaʻaongaʻi ha ʻaisi kaukau pe ʻaisi momoa ke mokomoko lolotonga e sonication. Tuku hoʻo (ngaahi) foʻi tiupi lolotonga e sonication ki ha falekaukau ʻi he ʻaisi. Ki he VialTweeter, fakaʻaongaʻi ha laulau mamaha ʻoku fonu ʻi he ʻaisi momoa pea tuku ʻa e VialTweeter ʻi he ʻaisi momoa koeʻuhi ke lava ʻo molia vave ʻa e vela.

ʻOku fakaʻaongaʻi ʻe he kau fakatau fakaemamani lahi ʻa e Hielscher fekumi-ultrasonicators pea pehe ki he sipinga fakavahaʻa-sonication ngaahi ʻiuniti VialTweeter mo UIP400MTP ki heʻenau sipinga teuteu fakaʻaho ʻi he founga totonu, biochemical, fakafaitoʻo mo kiliniki laboratories. Ko e polokalama fakakomipiuta poto mo e mafana ʻo e ʻea ʻoku ne puleʻi ʻa e Hielscher processors, ʻoku reliably mapuleʻi ʻa e mafana mo e sipinga ʻo e ʻea vela ʻoku ne fakaʻehiʻehi mei ai. ʻOku hanga ʻe ultrasonic sipinga ʻo e teuteu mo Hielscher ultrasonic ngaahi founga fakaleleiʻi ʻo fakahaofi ha ngaahi ola falalaʻanga mo reproducible!

Fetu'utaki mai kiate kimautolu! / Kole kiate kitautolu!

Kole ki ha fakamatala lahi ange

Kataki 'o ngaue 'aki e foomu 'i lalo ke kole ha toe fakamatala ki he ultrasonic processors, faka'aonga'i mo e totongi. Te mau fiefia ke alea'i e founga 'oku mo 'i ai mo ho'omou ngaahi fie ma'u ultrasonic he polokalama!









Kataki 'o fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


Ultrasonic high-shear homogenizers are used in lab, bench-top, pilot and industrial processing.

Hielscher Ultrasonics ʻa e ngaohiʻanga koloa maʻolunga ultrasonic homogenizers ki hono tuifio ʻo e ngaahi polokalama, fakamovetevetea, emulsification mo e toʻo hingoa ʻi he fale fakatotolo fakakemi, pailate mo e meʻafua ki he ngaahi meʻa fakapisinisi.

Ngaahi tohi/fakamo'oni fakafolofola



Ngaahi mo'oni'i me'a 'oku mahu'inga ke 'ilo'i

Faʻahinga ʻo ʻELISA

ʻOku lahi e ngaahi faʻahinga ʻo ʻELISA, ʻa ia ʻoku fakafaikehekeheʻi ʻe he tefitoʻi moʻoni ʻoku nau ngaue ʻaki. ʻOku ʻiloa kinautolu ko e fakahinohino fakahangatonu ʻa ʻELISA, ʻA ELISA, sanuisi ʻELISA, feʻauʻauhi mo ʻELISA, pea fulihi ʻa ʻELISA. ʻI lalo, ʻoku mau ʻoatu ha vakai fakalukufua ki he ngaahi faʻahinga kehekehe ʻo ʻELISA mo honau ngaahi tefitoʻi ʻulungaanga mo e faikehekehe.
ʻE lava ke lele ʻa ʻELISA ʻi ha founga qualitative pe quantitative. ʻOku ʻomi ʻe he ngaahi ola Qualitative ha ola lelei pe kovi, lolotonga ia ʻoku quantitative ʻa ʻELISA ko e density (OD) ʻo e sipinga ʻoku fakafehoanaki ia ki ha Pikoʻanga angamaheni, ʻa ia ʻoku angamaheni ʻaki hono fakahokohoko fakalanu

Tataki ʻa ʻELISA

Ko e fakahinohino fakahangatonu ko ia ʻa ʻELISA ko e founga faingofua taha ia ʻo ʻElisa, ʻa ia ʻoku fakaʻaongaʻi ai ha antibody Palaimeli enzyme pea ʻoku ʻikai fie maʻu ha maluEi maʻolunga. ʻOku haʻi fakahangatonu ʻa e enzyme Palaimeli ʻoku fakahingoa ko e antibody ki he taumuʻa, hange ko e antigen. ʻOku tanaki atu ʻa e founga buffered antigen ki he lelei takitaha ʻo ha peleti microtiter (meimei ko e ʻu lauʻi peleti lelei ʻe 96, ʻELISA-ʻu lauʻi peleti), ʻa ia adheres ki he tafaʻaki pelesitiki ʻo fakafou ʻi he ngaahi fengaueʻaki fekauʻaki mo e totongi. ʻI he taimi ʻoku fehokotaki ai ʻa e enzyme mo e antibody Palaimeli mo hono substrate, ʻoku ne ʻomi ha fakaʻilonga ʻe lava ke fua ʻo fakafou ʻi he spectrophotometer, fluorometer, pe luminometer.

ʻIkai fakahangatonu ʻa ʻELISA

ʻI he sivi ʻoku ʻikai fakahangatonu, ʻoku fakatou fie maʻu ai ha antibody Palaimeli mo ha antibody maʻolunga. Neongo ia, ʻoku fehangahangai ia mo e fakahinohino fakahangatonu ʻa ʻELISA, kae ʻikai ko e antibody Palaimeli, ka ʻoku fakahingoa ʻa e antibody hono ua ʻaki ha enzyme. ʻOku taʻofi ʻa e antigen ki he peleti lelei pea haʻi ʻe he antibody Palaimeli. ʻI he faifai pe pea haʻi ʻa e enzyme ua antibody ki he antibody Palaimeli. Fakaʻosi, ko e enzyme ʻoku fakafehokotaki ki he antibody hono ua mo ʻene substrate ke maʻu ha fakaʻilonga ʻoku lava ke fakatokangaʻi.

Sanuisi ʻELISA

Lolotonga e ngaahi sivi fakahangatonu mo ʻikai fakahangatonu ʻa ʻELISA, ʻoku taʻofi ʻa e antigen pea ʻave ia ki he fukahi peleti vaitupu, ʻi he sanuisi ko ʻELISA ʻoku taʻofi ʻa e antibody ki he tafaʻaki pelesitiki ʻo e peleti ʻa ʻELISA. ʻOku ʻiloa ʻa e maluEi maluEi ʻi he sanuisi ko ʻELISA ko e maluEi. Makehe mei he maluEi, ʻoku fie maʻu foki ke fai ha fakatokanga ki he maluEi ʻi he sanuisi ko ʻELISA. ʻOku kau ʻi he maluEi fakatokanga ha fakatokanga Palaimeli ʻoku ʻikai fakahingoa antibody mo ha fakatokanga maʻolunga enzyme antibody.
Stepwise, ʻoku haʻi ʻa e antigen ʻo e tokanga ki he maʻu antibody taʻofi ki he peleti. ʻI he taimi ko ia, ʻoku haʻi leva ʻa e tefitoʻi fakatokanga antibody ki he antigen. Hili ia, ʻoku haʻi ʻa e fakatokanga hono ua antibody ki he tefitoʻi fakatokanga antibody. ʻI he sitepu fakaʻosi, ʻoku toe ngaue ʻa e enzyme mo hono substrate ke maʻu ha fakaʻilonga ʻe lava ke fakatokangaʻi optically.

Feʻauʻauhi ʻa ʻELISA

Ko e feʻauʻauhi ʻa ʻELISA, ʻa ia ʻoku toe ʻiloa pe ko inhibition ʻELISA, ko e faʻahinga faingataʻa taha ia koeʻuhi he ʻoku kau ai hono fakaʻaongaʻi ʻo ha inhibitor antigen. ʻE lava ke fulihi ʻa e founga takitaha ʻe tolu, hangatonu, ʻikai fakahangatonu, mo e sanuisi ko ʻELISA, ki he founga feʻauʻauhi ʻa ʻELISA. ʻI he feʻauʻauhi ʻa ʻELISA, ko e antigen inhibitor mo e antigen ʻo e tokanga ke haʻi ki he antibody Palaimeli.
Ke feʻauʻauhi ʻa ʻELISA, ʻoku incubated ʻa e antibody ʻoku ʻikai ke ʻiloa ʻi he ʻao ʻo hono antigen, hange ko e sipinga. ʻOku tanaki atu leva ʻa e antibody/antigen complexes ko ʻeni ki ha antigen lelei.
ʻOku fufulu e peletí, ke toʻo e maluEi ʻoku ʻikai ke saí. ʻOku maʻu ʻe he feʻauʻauhi ʻa ʻELISA ʻa hono hingoa koeʻuhi ko e foʻi moʻoni ko ia ko e antigen lahi ange ʻoku ʻi he sipinga, ko e antigen antibody complexes ʻoku fokotuʻu. ʻOku ʻuhinga ʻeni, ʻoku siʻisiʻi ange ʻa e maluEi ʻoku lava ke maʻu ke haʻi ki he antigen ʻi he vaitupu pea kuo pau ke feʻauhi ʻa e antigens ki ha antibody ʻoku lava ke maʻu. ʻOku tanaki atu ha antibody hono ua, ʻo tatau mo e antibody Palaimeli. ʻOku fehokotaki ʻa e antibody hono ua ko ʻeni mo e enzyme. ʻI he taimi ʻoku tanaki atu ai ʻa e substrate, ʻoku ʻomi ʻe he toenga enzymes ha fakaʻilonga chromogenic pe maama tiupi.
ʻI he taimi ko ʻeni, ʻoku taʻofi ʻa e tali ke fakaʻehiʻehi mei he matolutolu ʻo e fakaʻilonga.
ʻOku kau ʻi he ngaahi naunau feʻauʻauhi ʻa ʻELISA ha antigen ʻoku fakafehokotaki enzyme kae ʻikai ko ha antibody ʻoku fakafehokotaki enzyme. ʻOku fakahingoa ʻa e antigen competes ki he palaimeli antibody ngaahi feituʻu ʻoku haʻi ʻaki ʻa e sipinga antigen (ʻikai fakahingoa). Ko e siʻisiʻi ange antigen ʻi he sipinga, ko e lahi ange ia hono fakahingoa antigen ʻoku tauhi ia ʻi he vaitupu pea malohi ange ʻa e fakaʻilonga.

Fulihi ʻa ʻELISA

ʻOku ʻikai fakaʻaongaʻi ʻe he tafaʻaki ʻe ʻELISA ʻa e ʻu lauʻi peleti lelei, ka ʻoku mavahe ia mei he antigens ʻoku tuku ʻi he lolo sivi. ʻOku fuatautau ʻe he sivi ʻa ʻELISA ʻa e lahi ʻo e antibody kuo haʻi ʻo fakafou ʻi he antigen. Naʻe fakatupulaki ia ke ʻiloʻi mo fakatotoloʻi ʻa e ivi sila ʻo e vailasi Vaitafe Naila mo e founga ʻoku lava ke maʻu ai ha maluEi pau ʻi he vailasi.

Enzymatic peni fakaʻilonga naʻe fakaʻaongaʻi ki he ʻELISA

ʻOku ʻoatu ʻe he lisi ʻi lalo ʻa e ngaahi fakaʻilonga enzymatic angamaheni taha ʻoku fakaʻaongaʻi ʻi he lea ʻa ʻELISA, ʻa ia ʻoku ne fakaʻata ʻa e ola ʻo e lea ke fakafuofuaʻi ʻi hono fakakakato.

  • ʻOku liliu ʻe OPD (o-phenylenediamine dihydrochloride) ʻa e ʻAmipa ke ʻiloʻi HRP (Horseradish Peroxidase), ʻa ia ʻoku faʻa fakaʻaongaʻi ko ha ivi conjugated.
  • TMB (3, 3′,5,5′-tetramethylbenzidine) ʻa e lanu pulu ʻi he taimi ʻoku fakaʻilo ai HRP pea liliu engeenga hili hono tanaki atu ʻo e sulfuric pe phosphoric ʻesiti.
  • ʻAPIKALE (2,2′-Azinobis [3-ethylbenzothiazoline-6-sulfonic ʻesiti]-diammonium masima) ʻi he taimi ʻoku fakaʻilo ai HRP.
  • ʻOku liliu lanu engeenga ʻa PNPP (p-Nitrophenyl Phosphate, Disodium masima) ʻi he taimi ʻoku fakaʻilo ai ʻalakalaine phosphatase.