ELISA nrog qhov tshwm sim tsis zoo?Ua raws li peb xaiv lub phaj Elisa tiag.
Xaiv lub phaj tsim ELISA yog thawj kauj ruam rau kev ua tiav ELISA kev sim, tab sis ntau tus neeg tsis quav ntsej qhov no. Thaum tib neeg ua qhov kev sim, lawv feem ntau tsuas yog xav txog qhov kev sim nws tus kheej, tab sis tsis paub tias cov khoom feem ntau muaj kev cuam tshuam zoo rau kev sim. Kev xaiv cov khoom tsim nyog yuav ua rau muaj kev vam meej ntawm kev sim. Peb feem ntau xav txog cov hauv qab no thaum ua qhov kev sim ELISA:
1.Cov duab hauv qab
Tam sim no, cov chaw tsim khoom tsim cov enzyme-labeled qhov dej raws li cov kab kev ywj pheej, thiab tsim kom muaj qhov kev ncua deb ntawm cov qhov dej kom lawv tuaj yeem sib cais ntawm nws tus kheej, yog li txo qis kev sib kis. Ntau hom Elisa daim hlau muaj cov duab sib txawv ntawm lub qhov dej hauv qab. Nov yog qee qhov sib xws hauv qab cov duab:
Flat hauv qab: Hauv qab yog kab rov tav, tseem hu ua F hauv qab. Lub teeb yuav tsis raug deflected hauv qab, uas tuaj yeem xa lub teeb mus rau qhov loj tshaj plaws. Siv rau kev sim uas xav tau ib puag ncig hauv qab rau kev pom lossis lwm yam laj thawj.
Round hauv qab: Kuj hu ua U-hauv qab, muab cov txiaj ntsig zoo tshaj plaws thiab kev ua haujlwm sib xyaw ua ke, tsim nyog rau cov ntawv thov uas yuav tsum tau kuaj cov sediments.
C-hauv qab: Nws tuaj yeem muab kev tu zoo ntawm lub tiaj tiaj hauv qab thiab lub hauv qab puag ncig, thaum sib txuas qhov zoo ntawm lub tiaj tiaj hauv qab.
Lub khob hauv qab: Kuj tseem hu ua V-hauv qab, nws yog qhov tsim nyog rau kev kuaj xyuas meej thiab khaws cov qauv micro rau kev pom zoo rov qab ntawm cov ntim me me.
2. Cov xim ntawm Elisa phaj
Muaj peb xim ntawm Elisa phaj: pob tshab, dub thiab dawb.
Feem ntau ntawm ELISA xaiv cov ntawv pob tshab los ua cov ntaub ntawv sim. Dawb thiab dub Elisa daim hlau feem ntau yog siv rau kev kuaj pom luminescence. Lub phaj dub ELISA nws tus kheej nqus lub teeb, yog li nws cov teeb liab tsis muaj zog dua li ntawm ELISA phaj dawb. Cov phaj dub Elisa feem ntau yog siv los kuaj pom lub teeb muaj zog, xws li kev kuaj pom fluorescence; ntawm qhov tsis sib xws, cov ntawv dawb Elisa tuaj yeem siv rau kev pom lub teeb pom kev tsis muaj zog, thiab feem ntau siv rau kev siv tshuaj lom neeg thiab cov xim substrate (xws li dual luciferase). reporter gene analysis).
3. Cov khoom ntawm Elisa phaj
Cov khoom siv feem ntau yog polyethylene (PE), polypropylene (PP), polystyrene (PS), polyvinyl chloride (PVC) thiab polycarbonate (PC).
Cov ntaub ntawv siv dav tshaj plaws hauv ELSIA yog polystyrene thiab polyvinyl chloride. Polyvinyl chloride soft board yog nyias, tuaj yeem txiav, thiab tus nqi qis. Qhov tsis zoo yog qhov ua tiav tsis zoo li lub rooj tsavxwm polystyrene, thiab hauv qab ntawm lub qhov tsis tiaj li polystyrene. Tab sis tus nqi keeb kwm yav dhau los kuj yuav nce ntxiv. Feem ntau, ion grafting kev kho mob yog nqa tawm nyob rau saum npoo ntawm ELISA phaj, thiab active functional pab pawg xws li aldehyde pawg, amino pawg, thiab epoxy pawg yog nkag mus rau hauv lub polymer nto los txhim kho kev ua tau zoo ntawm lub substrate nto.
4. sib txawv binding mechanisms
Kev sib khi zoo ntawm cov khoom siv txheej rau hauv qab ntawm lub phaj yog ib kauj ruam tseem ceeb hauv ELISA. Yog li ntawd, cov yam ntxwv ntawm cov yam ntxwv thiab cov khoom siv tshuaj lom neeg ntawm cov khoom yuav tsum tau coated yuav tsum tau ua tib zoo txheeb xyuas ua ntej qhov kev sim los xaiv ib lub phaj Elisa. Feem ntau hais lus, muaj ntau txoj hauv kev los khi cov khoom siv rau hauv qab ntawm lub phaj: passive adsorption, covalent binding thiab affinity capture.
Passive Adsorption: Cov txheej txheem ntawm kev nqus adsorption yog qhov nyuaj, thiab kev sib txuas ntawm cov khoom siv txheej thiab hauv qab ntawm cov phaj feem ntau yog nyob ntawm cov rog intermolecular (van der Waals force) thiab hydrogen bonds. Nyob rau hauv feem ntau ELISAs, kev khi ntawm cov tshuaj txheej rau hauv qab ntawm lub phaj yog ua tiav los ntawm passive adsorption. Feem ntau, kev khi ntawm nruab nrab thiab loj molecular hnyav antigens (los yog cov tshuaj tiv thaiv) thiab hauv qab ntawm lub phaj yog ua tiav los ntawm passive adsorption. Cov yam ntxwv hydrophobic/hydrophilic ntawm lub hauv qab ntawm lub phaj feem ntau xav txog thaum xaiv ib lub phaj Elisa. Cov lus hauv qab no teev cov peev txheej ntawm daim ntawv thov thiab siv cov khoom sib txawv hydrophilic substrate:
hydrophilic | Kev khi ua ntej | Qhov tseem ceeb siv |
- | Molecules uas muaj cov cheeb tsam tseem ceeb hydrophobic, xws li: lipids, lipoproteins, cov proteins loj | Antigen ELISA, FlaA, LIA |
ntxiv | Biomolecules nrog hydrophilic / hydrophobic zog xws li: nruab nrab mus rau loj proteins, immunoglobulins, albumin | Ob chav Antibody Sandwich ELISA, Antigen ELISA |
plus ntxiv | Biomolecules nrog hydrophilic / hydrophobic zog xws li: me me mus rau loj proteins, immunoglobulins, albumin, LPS, phosphorylated proteins, glycoproteins | Ob chav antibody ELISA, FIA, LIA |
plus ntxiv | Glycoproteins, polar lipid molecules, phospholipids, cardiolipids | Antigen ELISA |
Affinity Capture: Affinity capture yog raws li qhov tshwj xeeb khi ntawm cov ntawv sau macromolecules rau lawv cov receptors sib raug. Tagged macromolecules tuaj yeem tau txais los ntawm kev sib txuas tshuaj lom neeg lossis los ntawm genetic engineering.
Nta ntawm Affinity Capture yog:
1.high specification
2.Can txhim kho lub teeb liab-rau-suab nrov piv ntawm kev ntsuas ntsuas
Qhov zoo tshaj plaws hauv qab qhov chaw yog: streptavidin (streptavidin) coated nto; nickel chelate (Nickel Chelate) coated nto thiab glutathione (Glutathione, GSH) coated nto.
3. Streptavidin nto: Lub siab affinity thiab tshwj xeeb ntawm streptavidin thiab biotin yog siv los paub txog kev khi ntawm cov phaj hauv qab rau cov protein coated.
4. Nickel chelate nto: Nickel chelate yog txuas rau lub polymer nto los khi fusion proteins nrog Nws (histidine) cov cim npe.
5. Glutathione nto: Siv cov enzyme thiab substrate tshwj xeeb ntawm glutathione thiab glutathione sulfhydryltransferase (GST) kom ua tiav kev khi ntawm cov phaj hauv qab rau cov txheej txheem protein.
Vim li cas thiaj xaiv BKMAM Elisa phaj?
Ua los ntawm 100% cov khoom siv polystyrene,
Haum rau adherent cell culture,
Pav ca puag, txawm thickness thiab uniform loj rau cov tshuaj tiv thaiv ncej,
Sterile nrog tus kheej pob,
High quality nrog Hoobkas nqi.
