Sars-cov-2 kusarerekera antibody yekuona kit (Elisa)
【Nheyo】
Iyo inochinjisa-cov-2 kusarerekera Anti
Kushandisa RECFED Receptor Binding Domain (RBD), protein kubva kuVIRIRA Spike (s) protein uye sero rekutambira
Receptor Ace2, muedzo uyu wakagadzirirwa kutevedzera hutachiona hwekutakura hutachiona.
Calibrators, mhando dzemhando yepamusoro, uye serum kana samples plasma dzinosanganiswa pamwe chete mukudhura
Buffer ine Hace2-hrp conjugate yakagadziriswa mumabhuru madiki. Ipapo musanganiswa unoendeswa mukati
iyo microplate matsime ane demaliblised sars-cov-2 rbd chidimbu (rbd) ye
kuhukura. Munguva yemaminitsi makumi matatu-yemindu, iyo rbd chaiyo antibody mune calibrators, qc uye
Sampuli dzinokwikwidza neHACE2-HRP yekunzwisisa chairo iyo RBD yakabatanidzwa mumatsime. Mushure
Iyo incubation, iyo matsime anogezwa ka4 nguva yekubvisa Unbound Hace2-hrp conjugate. Mhinduro ye
TMB inowedzerwa uye yakavezwa kwemaminitsi makumi maviri pamhepo tembiricha, zvichikonzera mukuvandudzwa kwe
ruvara rwebhuruu. Iyo yekuvandudza kwemavara kunomira pamwe nekuwedzera kwe 1n HCL, uye iyo yekubata ndeye
yakayerwa spectrophotometricomselly at 450 nm. Kukura kwemavara kwakaumbwa kunoenderana ne
Hurefu hwe enzyme riripo, uye hune hukama nehuwandu hwehuwandu hwakafuratirwa nenzira imwe chete.
Kuburikidza nekuenzanisa neyakagadziriswa curve yakaumbwa neakapihwa calibrators, kusunganidzwa kwe
kusarerekera antibodies mune iyo isingazivikanwe sample inobva yaverengerwa.
【Zvigadzirwa zvinodiwa asi zvisina kupihwa】
1. Yakadzikira kana kuora mvura
2. Pipisi pipettes: 10μL, 100μl, 200μL uye 1 ml
3. Kuraswa Pipette Matipi
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5. Bepa rinobata
6. Giraira pepa
7. Vortex musanganiswa kana kuenzana
【Kuunganidza kuunganidza uye kuchengetedza】
1. Serum uye plasma samples yakaunganidzwa mumabhuru ane K2-edta inogona kushandiswa kit iyi.
2.
Fungidziro dzakabatwa kwenguva yakareba
Dzivisa kudzokororwazve-thaw matatya.
Protocol
【Reagent Kugadzirira】
1. Zvese Reagents inofanira kubviswa kubva kufiriji uye inobvumidzwa kudzoka kumhepo tembiricha usati washandisa
(20 ° kusvika 25 ° C). Sevha vese reagents mufiriji nekukurumidza mushure mekushandisa.
2. Zvese sampuli uye kutonga kunofanirwa kuvezwa kusati kwashandiswa.
3. Hace2-HRP Solution Kugadzirira: Dilute Hace2-HRP Yakatarisana na1: 51 Diliation Ratio neDulition
Buffer. Semuenzaniso, Dilute 100 μL yeHACE2-HRP inotarisa ne 5.0ml yeHRP Dailition Buffer to
gadzira hace2-hrp solution.
4. 1 × Wash Solution Kugadziriswa: Dilute iyo 20
Vhoriyamu Ratoo ye1:19. Semuenzaniso, Dilute 20 ml ye20 × SADE SUPEL ine 380 mL ye deionized kana
Mvura yakadzurwa kugadzira 400 ml ye1 × yekushambidza solution.
【Kuongorora maitiro】
1. Mune zvikamu zvakasiyana, aliquot 120μL yeiyo yakagadzirirwa yeHALE2-HRP Solution.
2. Wedzera 6 μL ye calibrators, samples isingazivikanwe, mhando dzekudzora mune yega thubhu uye sanganisa zvakanaka.
3. Chinjana 100μL yemusanganiswa wega wega akagadzirirwa mudanho 2 kuita zvinoenderana nemicrate microplate matsime maringe
kufanorongerwa kuongororwa kweyedzo.
3. Kufukidza ndiro neine mapani uye incubate pa37 ° C kwemaminetsi makumi matatu.
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5. Tinya ndiro papepa repepa kuti ubvise zvakasara mvura mumatsime mushure mekugeza matanho.
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7. Wedzera 50 μl yemhinduro yekumisa kune yega yega yetsime kumira kuita.
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inokurudzirwa pakubata kwakakwirira kwekuita).