Brucella antibdy Elisa Kit

Chifanizo

 

Njira Zoyeserera & Kutanthauzira

 

Gawo 1 Njira Yoyeserera

1.1 Sambani njira yokonzekera

WOrkiction ndende wkusamba madzi:Musanagwiritse ntchito, yankho losambitsa liyenera kubwezeretsedwanso kutentha (20-25 kuchuluka kwake) ndikugwedeza mokwanira kuti ipange yunifolomu. Kenako muchepetse 20 maulendo 20

1.2 Kukonzekera kwachiwiri

Magazi a nyama ya nyama ya nkhosa yamkati, ya centrift pa 4000rpm kwa mphindi 10, ndipo supernanatant adasonkhanitsidwa. Seramu imatha kuphatikizidwa mwachilengedwe popanda hemolysis.

Kukula kwa zitsanzo ndikokulira, kuti awonetsetse kuti nthawi yoyeserera yogwirizana ndi yogwirizana. Sakanizani zitsanzo za seramu ndi njira yogwiritsira ntchito mankhwala a antibody pa 1: 1 mu sheramu ya dilchetion (70ul sheramu (1) serram sterum (+ 70}} ul antibody ntchito yankho).

1.3 Njira Yoyeserera

①Take microplate ya enzyme (malinga ndi kuchuluka kwa zitsanzo, zitha kusokonezedwa ndikugwiritsa ntchito zitsime ziwiri) Pang'onopang'ono gwedezani chitsimechi pachitsime, pogwiritsa ntchito filimu yosindikizira mbale kuti muphimbe mbaleyo, ndikupindika pa 37 digiri kwa mphindi 30.

②opn filimu yosindikiza ya mbale, sinthani yankho mu mbaleyo bwino, onjezani 300μl yogwira ntchito kusamba madzi pachitsime chilichonse pachitsime. Bwerezani masitepe omwe ali pamwambapa kasanu, ndipo pamapeto pake pakhala pepala lopanda tanthauzo loyamwa.

③DIliated enzyme conjugate:Gawani enzyme conjugate (50 × 2 (mwachitsanzo: 20uln

④add 100μme conjuguate pachitsime chilichonse, kuphimba mbaleyo, ndikuphatikizira pa 37 digiri kwa mphindi 30.

Vuto la Enzyme Enzyme, opaleshoni inayake ndi yofanana ndi yomwe ili pamwambapa ②.

⑥add 100μl njira yothetsera vuto lililonse, kuphimba mbaleyo ndikuphatikizira kwa mphindi 10 pa digiri 37 kutali ndi kuwala.

⑦add 50μl siyani yankho la chitsime aliyense, ndikuzindikira zotsatira zosakwana mphindi 5.

 

Gawo 2 Zotsatira

 

Mtengo wa chitsime aliyense wa OD45 umawerengedwa pa Micreplate ya Microphe ya 96.

2.1

 

Mtengo wapamwamba wa OD450nm wowongolera anali <0.35 ndi kuchuluka kwa odetsa olakwika / kuwongolera bwino kunali kwakukulu kuposa kapena kofanana ndi 3.5, zoyeserera zinali zovomerezeka. Kupanda kutero, mayeso amayenera kubwerezedwa.

Njira ya 2.2 njira

Kwa sample iliyonse, kuwerengetsa pi

2.3 Chotsatira

Pamene pi <20%, imaweruzidwa ngati zoipa.

Pamene pi> 30%, imaweruzidwa kuti nditsimikizire.

20% yocheperako kapena yofanana ndi pi yocheperako kapena yofanana ndi 30%, imaweruzidwa monga kukayikira, ndikuwonetsa kubwereza zitsanzo sabata imodzi.

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