I-SARS-cov-2 engathethekiyo i-Anti Kiking Kit (Elisa)
】Umgaqo】
I-SARS-2 engathethekiyo i-antiction ikhithi ye-antids isekwe kwindlela yokhuphiswano lwe-ELISA.
Ukusebenzisa i-Captor ehlanjululwe ngu-Deiain (i-RBD), iprotein evela kwi-chikein spike (s) neprotheyini kunye neseli ye-vist
Ukufumana i-ACE2, olu vavanyo lwenzelwe ukulingisa intsholongwane yokungathathi cala.
Ababhali, ulawulo lomgangatho, kunye ne-serum okanye iisampulu zeplaum zidityaniswe kakuhle kwi-dilution
I-Buffer equkethe i-hace2-hrp ye-audioote engacacanga kwiityhubhu ezincinci. Emva koko umxube udlulisiwe
I-microplate yequla eline-SassIllIll engafakwanga kwi-SARS-cov-2 i-RBD (i-RBD) ye
I-incubation. Ngexesha lokufakwa kwemizuzu engama-30, i-rbd antiyd antis kwi calibrators, i-QC kunye
Iisampulu ziya kukhuphisana ne-hace2-hrp ye-RBD engafakwanga i-RBD equleni. Emva
Ukufakwa, amaqula ahlanjwe amaxesha ama-4 ukususa i-hace2-hrp conjugate. Isisombululo se
I-RB iyongezwa kwaye ifakwe imizuzu engama-20 kubushushu begumbi, kukhokelela kuphuhliso lwe
umbala ohlaza okwesibhakabhaka. Uphuhliso lwemibala lumiswe ngokongezwa kwe-1n hcl, kunye ne-absorby
ukulinganiswa i-spectrametrictricctrict nge-450 NM. Ubunzulu bombala owenziwe buhambelana ne
Isixa se-enzyme ekhoyo, kwaye sinxulumene nenani lemigangatho eyathathwa ngendlela efanayo.
Ngokubhekiswa kwi-calibrication curve eyenziwa ngababhayili abanikezelweyo, uxinzelelo lwe
Ukungathathi cala kwii-antibodies kwisampulu engaziwayo ibalwa.
】Izinto ezifunekayo kodwa azibonelelwanga】
I-1. Amanzi anelisiweyo
2. I-pipettes echanekileyo: I-10μl, 100 μl, 200μl kunye ne-1 ml
I-3. Iingcebiso zePipette ezilahlayo
I-4. Umfundi we-mncinci dafu enakho ukufundela i-abseys kwi-450nm.
I-5. Iphepha le-Absorber
Iphepha le-6. Iphepha legrafu
I-7. Umxube we-vrtex okanye ilingana
】Ukuqokelelwa kwengqokelela kunye nokugcina】
I-1. I-serum kunye neesampulu zeplasma eziqokelelwe kwiityhubhu eziqulathwe k2-Edta zingasetyenziselwa le kit.
2. Iisampuli kufuneka zibanjiwe kwaye zinokugcinwa ukuya kuthi ga kwiiyure ezingama-48 nge-2 ° C-8 ° C - 8 ° C ngaphambi kokuthambisa.
I-capimens ibambe ixesha elide (ukuya kuthi ga kwiinyanga ezi-6) kufuneka ikhule kuphela nge-1 ° C ngaphambi kwe-ASESY.
Khusela imijikelezo ephindaphindiweyo ye-freeze-thaw.
Iprotocol
】Ukulungiswa kwe-regent】
1. Zonke izinto kufuneka zikhutshwe kwifriji kwaye zivunyelwe ukubuyela kubushushu begumbi ngaphambi kokusebenzisa
(20 ° ukuya kwi-25 ° C). Gcina zonke iiRegeds kwifriji yefriji ngokukhawuleza emva kokusetyenziswa.
2. Zonke iisampulu kunye nolawulo kufuneka zibethelelwe ngaphambi kokusetyenziswa.
I-3. I-Hace2-HRP yokulungiselela: I-Hacele Hace2-HRP igxile ku-1: 51 Umyinge we-chalution nge-dilution
Buffer. Umzekelo, i-dilute 100 μl ye-hace2-hrp igxila nge-5.0ml ye-HRP Dilution buffer
Yenza isisombululo se-hace2-hrp.
I-4. 1 × 1
Umyinge we-1:19. Umzekelo, i-dilute 20 ml ye-20 × 20 ye-380 ml ye-deoed okanye
Amanzi awotshiweyo enze i-400 ml ye-1
】Inkqubo yovavanyo】
1. Kwiityhubhu ezahlukeneyo, iAliquot 120μl yesisombululo esilungiselelwe i-HRP2-HRP.
2. Yongeza i-6 μl yeCalibrators, iisampulu ezingaziwayo, ulawulo lomgangatho kwityhubhu nganye kwaye uxube kakuhle.
3. Dlulisa i-100μl yomxube ngamnye olungiselelwe inyathelo 2 kwi-microplate efanayo
ukucwangciswa kwangaphambili kovavanyo.
3. Gquma iplate kunye nePokeri yePlayiti kwaye ubumbele kwi-37 ° C kangangemizuzu engama-30.
4. Susa i-Plate Toaller kwaye uhlambe ipleyiti malunga ne-300 μm ye-1 μ ye-1 ye-1 ye-1 μ ngesisombululo sendalo esiline.
5. Cofa ipleyiti kwitawuli yephepha ukususa ulwelo lwentsalela kwiqula emva kokuhlamba.
6. Yongeza i-100 μm yesisombululo se-TMB nganye ye-RMB nganye kwaye ifake iplate ebumnyameni nge-20 ukuya kwengama-25 ° C kangangemizuzu engama-20.
7. Yongeza i-50 μl yesisombululo sokumisa kwi-pin nganye ukuba iyeke ukuphendula.
8. Funda i-Absorbed kumfundi weMicroplate nge-450 NM ngaphakathi kwemizuzu eli-10 (i-630nm njenge
Kuyacetyiswa ukuba kwenziwe umsebenzi ophezulu wokuchaneka).