I-SARS-Cov-2 NOTRALIC
【Isimiso】
I-SARS-Cov-2 yokususa i-antilody dection Kit isuselwa kwindlela yokuncintisana e-Elisa Methodegy.
Kusetshenziswa isizinda esihlanzayo esibophayo (i-RBD), amaprotheni avela kumaprotheni we-viral spike (s) neseli lokusingathwa
I-Receptor ACE2, lokhu kuhlola kwenzelwe ukulingisa ukusebenzisana kokungathathi hlangothi kwegciwane.
AmaCalibrators, izilawuli zekhwalithi, kanye namasampula e-serum noma ama-plasma ahlanganiswe ngokuhlatshelwa kahle nge-dilution
I-Buffer equkethe i-hace2-HRP conjugate i-Aliquited kumashubhu amancane. Lapho-ke izingxube zidluliselwa ngaphakathi
ama-microplate Wells aqukethe ama-SARS ahlangene ahlaziya kabusha-cov-2 rbd (RBD) we
Ukufakwa. Ngesikhathi sokufakwa umzuzu wemizuzu engama-30, i-antibody ethize ye-RBD kuma-calibrators, QC futhi
Amasampula azoncintisana ne-Hace2-HRP ngokubopha i-RBD engasebenzi emithonjeni. Kamuva
Ukufakwa, imithombo iyagezwa amahlandla ama-4 ukususa umhlanganisi we-hace2-HRP ongenampilo. Isixazululo se
I-TMB bese ingezwa futhi ifakwe imizuzu engama-20 emazingeni okushisa asekamelweni, okuholela ekuthuthukisweni kwe
Umbala oluhlaza okwesibhakabhaka. Ukuthuthukiswa kombala kuyamiswa ngokufakwa kwe-1n hcl, futhi ukumunca
ulinganise i-spectrophotometrically ku-450 nm. Ukuqina kombala okwakhiwe kulingana ne
Inani lama-enzyme akhona, futhi ahlobene kakhulu nenani lamazinga atholakalayo ngendlela efanayo.
Ngokuqhathanisa namajika okulinganisa akhiwa ama-calibrators anikezwe, ukugxila kwe
Ama-antibodies angathathi hlangothi kusampula engaziwa ayakubalwa.
【Izinto ezidingekayo kodwa azinikeziwe】
1
2. I-Pipette Eqondile: I-10μL, 100μL, 200μL ne-1 ml
3. Izeluleko ze-Pipette ezilambile
4. I-Microplate Reader enekhono lokufunda ngo-450nm.
I-5. Iphepha elimunca
6. Iphepha legrafu
7. I-Vortex Mixer noma efanayo
【Ukuqoqwa kweqoqo nesitoreji】
1. Amasampula e-serum ne-plasma aqoqwe amashubhu aqukethe i-K2-Edta angasetshenziselwa leli khithi.
2
Izinhlobo ezenzelwe isikhathi eside (kuze kube yizinyanga ezi-6) kufanele zifakwe iqhwa kuphela kanye -20 ° C ngaphambi kokufa.
Gwema imijikelezo ephindaphindwayo ye-freeze-thaw.
Umthetho olandelisayo
【Ukulungiswa kweReagent】
1. Zonke iziphendulweni kufanele zikhishwe esiqandeni futhi zivunyelwe ukubuyela ekushiseni kwegumbi ngaphambi kokusebenzisa
(20 ° kuya ku-25 ° C). Gcina wonke ama-reagents esiqandisini ngokushesha ngemuva kokusetshenziswa.
2. Wonke amasampula nezilawuli kufanele zifakwe ngaphansi kokusetshenziswa.
I-3. I-HaCE2-HRP Solution Ukulungiselela: Dilite Hace2-HRP Gxila ku-1: 51 Dilution Ratio nge-dilution
Buffer. Isibonelo, jw μl ye-HACE2-HRP Gxila nge-5.0ML ye-HRP Dilution Buffer to
yenza ikhambi le-hace2-HRP.
4. 1
isilinganiso sevolumu sango-1: 19. Ngokwesibonelo, dilite 20 ml yesixazululo esingu-20 geza esingu-380 ml se-deionized noma
Amanzi ahlanganisiwe ukwenza i-400 ml yesisombululo sokugeza esingu-1 ×.
【Inqubo yokuhlola】
1. Kumashubhu ahlukene, i-aliquot 120μL yesisombululo esilungiselelwe i-Hace2-HRP esilungiselelwe.
2. Faka ama-6 μl ama-calibrators, amasampula angaziwa, izilawuli zekhwalithi kushubhu ngalinye bese uxuba kahle.
3
ukumiswa kokuhlolwa kokuhlolwa.
3. Mboza ipuleti nge-Plate Sealer futhi ufaka ku-37 ° C imizuzu engama-30.
4. Susa umthengisi wepuleti bese ugeza ipuleti nge-300 μL yesisombululo sokugeza esingu-1 × ngamahlandla amane.
I-5. Thepha ipuleti kuthawula lephepha ukuze ususe uketshezi oluyinsalela emithonjeni ngemuva kokugeza izinyathelo.
6. Faka isixazululo se-100
7 Faka i-50 μL yesisombululo sokuma kumthombo ngamunye ukumisa ukusabela.
8. Funda ukumunca ku-Microplate Reader ngo-450 nm kungakapheli imizuzu eyi-10 (630nm njenge-accessory
Kunconyelwe ukusebenza okuphezulu kwe-PROCION).