SARS-COV-2 Kubogamiye AntiBy Gutahura Kit (Elisa)
【Ihame】
Ibihe bya Sars-Cute-2 Kubogamiye AntiBy Gutahura Kit ishingiye kumarushanwa ya Elisa Methodedoology.
Gukoresha Receptor Ihuza domaine (RBD), Proteine kuva prokisi ya virusi hamwe na poroteyine na selile
Kwakira Ace2, iki kizamini cyagenewe kwigana imikoranire ya virusi.
Calibrators, Igenzura ryiza, na serumu cyangwa ingero za plasma zivanze kugiti cye zivanze
Buffer irimo Hace2-HRP ihuriweho na bilike mumiyoboro mito. Noneho imvange yimurirwa muri
Microplate Iriba irimo Scombinant Stombinant Sars-Cov-2 RBD (RBD) kuri
incubation. Mugihe cyiminota 30, Antibody yihariye muri calibrators, qc na
Ingero zizarushaho guhangana na Hace2-HRP kugirango ishyireho RBD idahumuriza mumariba. Nyuma
Incubations, amariba yogejwe inshuro 4 kugirango ukureho Hace2-HRP conjugate. Igisubizo cya
TMB yongeyeho kandi irashingwa muminota 20 mubushyuhe bwicyumba, bikaviramo iterambere rya a
ibara ry'ubururu. Iterambere ryamabara rirahagarara hamwe na 1n hcl, kandi absorbance ni
gupimwa neza kuri 450 Nm. Ubukana bwibara ryakozwe nigereranijwe kuri
Umubare wa enzyme uhari, kandi ufitanye isano muburyo butandukanye nubuziranenge bwamahame asaba muburyo bumwe.
Unyuze mugereranya n'umurongo wa kalibrasi wakozwe na kalibratory yatanzwe, kwibanda kuri
Kudahindura Antibodies mumitsindi itazwi noneho ibarwa.
【Ibikoresho birakenewe ariko ntibitangwa】
1. Amazi yandujwe cyangwa igicucu
2. Pipettes Precisite: 10μl, 100μl, 200μl na 1 ml
3. Inama za pipette
4. Umusomyi wa Microplate ashoboye gusoma kwinjiza saa yitanu.
5. Impapuro zinjira
6. Impapuro
7. Vortex ivanze cyangwa bihwanye
【Icyegeranyo cya Specimen】
1.
2. Ingero zigomba gutegurwa kandi zirashobora kubikwa kumasaha 48 kuri 2 ° C - 8 ° C mbere yo guhishurira.
Ibyingenzi byabaye igihe kirekire (kugeza kumezi 6) bigomba gukonjeshwa rimwe gusa kuri -20 ° C mbere yo gusaba.
Irinde gusubiramo gukonjesha-thaw cycle.
Protocole
【Gutegura reagent】
1. Ibishishwa byose bigomba gukurwa muri firigo kandi byemererwa gusubira mubushyuhe bwicyumba mbere yo gukoresha
(20 ° kugeza 25 ° C). Uzigame reagents zose muri firigo vuba nyuma yo kuyikoresha.
2. Ingero zose nubugenzuzi bigomba gutorwa mbere yo gukoresha.
3. Hace2-HRP Igisubizo cyo Gutegura: Dilute Hace2-HRP Yibanze kuri 1: 51 Ikigereranyo cyo Kwirukana hamwe no Kwirukana
Buffer. Kurugero, dilute 100 μl ya Hace2-HRP yibanda kuri 5.0ml ya HRP yohereza buffer kuri
kora igihome2 cya HRP.
4.
igipimo cy'ijwi rya 1:19. Kurugero, dilute 20 ml ya 20 × Karaba hamwe na ml 380 ya ml ya deionized cyangwa
Amazi yatowe kugirango akore 400 mL ya 1 × yoza igisubizo.
【Uburyo bw'ikizamini】
1. Mubyiciro bitandukanye, aliquot 120μl byigihuru cyateguwe2-HRP.
2. Ongeraho 6 μl ya calibrators, ingero zitazwi, igenzura ubuziranenge muri buri gituba hanyuma uvange neza.
3. Kohereza 100μl ya buri mvange yateguwe mu ntambwe ya 2 muri microplate licplate ihuye ukurikije
Kuri Iboneza ryibizamini.
3. Gupfuka isahani hamwe na papa hamwe na intera kuri 37 ° C muminota 30.
4. Kuraho umupira wamasahani hanyuma ukarabe isahani hafi 300 μl ya 1 × koza igisubizo kuri ererforfor inshuro enye.
5. Kanda icyapa kumpapuro kugirango ukureho amazi asigaye mumariba nyuma yo gukaraba intambwe.
6. Ongeraho 100 μl ya TM TM TMS
7. Ongeraho 50 μl yo guhagarika buri kintu kugirango uhagarike reaction.
8. Soma ibyuma muri microplate umusomyi wa 450 nm muminota 10 (630nm nkibikoresho ni
Basabwe kubikorwa byikirenga).