Tiibishada Bovine Antibody Elisa Kit

Tilmaamid

BTB Ab Elisa

Isticmaalka la damacsan yahayXirmada ogaanshaha ELISA ee ka hortagga qaaxada Bovine waxaa loogu talagalay go'aaminta qaaxada lo'da

antibody ee serum bovine ee Mycobacterium bovis in vitro.

Baaritaanka waxaa loo isticmaalaa in lagu qiimeeyo caabuqa qaaxada lo'da. MPB70-MPB83 chimera recombinant ayaa lagu dabaqay tijaabadan.

Tijaabada: serum

Alaabta Loo Baahan Yahay Laakiin Aan La Bixin:

Biyo sifaysan

Pipettor

96-si fiican u akhriye Microplate (450nm)

Saacadaha

Centrifuge (4000rpm)

Mashiinka dharka

Foornada (37 darajo)

Habka Imtixaanka & Fasiraadda

TALLAABADA 1Habka Imtixaanka

1.1 Diyaarinta Xalka Dhaq

Wfiirsashada orking wdareere dambas ah:Ka hor inta aan la isticmaalin, xalinta dhaqidda waa in lagu soo celiyaa heerkulka qolka (20-25 digrii) oo si siman loo ruxayaa si ay u noqoto mid labbis ah. Dabadeed ku qas 20 jeer xalalka dhaqidda ee xoogga leh oo leh biyo nadiif ah (tusaale ahaan, 5ml xal dhaqid oo xoog leh ayaa lagu daraa 95ml biyo nadiif ah), si fiican isku qas, dareeraha dhaqidda ee la qasi karo waxaa lagu kaydin karaa 2-8 digrii 7 maalmood.

1.2 Tusaalaha Diyaarinta

① Dhiigga xoolaha oo dhan ayaa la qaaday, iyadoo la saaray 4000rpm 10 daqiiqo, waxaana la ururiyay subagga sare. Serum sidoo kale si dabiici ah ayaa loo xinjiraysan karaa iyada oo aan la helin dhiig-baxa.

② Ku qas muunadda serumka 1:100 (ku dar 2ul serum muunad 198ul muunad dareere ah) oo si fiican isku qas.

Fiiro gaar ah: Kontaroolada xun iyo kuwa togan looma baahna in la qaso.

1.3 Diyaarinta isku dhafka Enzyme

Ku milmi isku-xidhka enzyme 1:200 (ku dar 5ul enzyme conjugate (x100) ilaa 995ul muunad dareere ah) oo si fiican isku qas.

1.4 Habka Imtixaanka

Qaado calaamadda enzyme Microplate (sida ku cad tirada shaybaarrada, waa la kala diri karaa oo la isticmaali karaa dhowr jeer), ku dar 100ul ee serum-ka la qasi jiray calaamadda enzyme Microplate, dhig laba ceel midkiiba muunado taban iyo mid togan, oo ku dar 100ul mid kasta oo loogu talagalay xakamaynta taban iyo kuwa togan ee falcelinta si fiican u kala horreeyaan. Si tartiib ah u rux muunada ceelka, adigoo isticmaalaya filimka saxanka daboolaya si aad u daboosho saxanka, oo ku dabool 37 darajo 30 daqiiqo.

② Fur filimka saxanka, ka rux xalka saxanka si fiican, ku dar 300µl oo dareere dhaqid ah oo shaqaynaya ceel kasta, ka dibna tuur dareeraha ceelka. Ku celi tillaabooyinka kore 5 jeer, oo ugu dambeyntii ku qallaji warqad nadiif ah oo nuugta.

③Ku dar 100µl of Enzyme Conjugate ceel kasta, dabool saxanka, oo ku duri 37 darajo 45 daqiiqo.

④ Fur filimka saxanka, ka rux xalka saxanka si fiican, ku dar 300µl oo dareere dhaqid ah oo shaqaynaya ceel kasta, ka dibna tuur dareeraha ceelka. Ku celi tillaabooyinka kore 5 jeer, oo ugu dambeyntii ku qallaji warqad nadiif ah oo nuugta.

⑤Ku dar 100µl xal substrate ceel kasta, dabool saxanka oo ku dabool 10 daqiiqo meel 37 digrii ka fog iftiinka.

⑥Ku dar 50µl xal joogsi ah ceel kasta, oo go'aami natiijada 5 daqiiqo gudahood.

TALLAABADA 2 Fasiraadda Natiijooyinka

Qiimaha OD450 ee ceel kasta waxaa lagu akhriyay 96-well microplate akhristaha.

2.1 Xaqiijinta

Celceliska qiimaha OD450nm ee xakamaynta taban wuxuu ahaa<0.5 and the average OD450nm value of positive control was ≥1, the experimental results were valid. Otherwise, the test should be repeated.

2.2 Habka Xisaabinta

Muunad kasta, u xisaabi S/P sida soo socota adoo isticmaalaya muunada la saxay iyo qiimayaasha xakamaynta:

2.3 Fasiraadda Natiijooyinka

Marka S/P<0.25, it is judged as negative.

Marka S/P ka weyn yahay ama la mid yahay 0.30, waxa loo qiimeeyaa mid togan.

Marka 0.25 ka yar ama la mid yahay S/P<0.30, it is judged as suspicious, and suggest to repeat sampling and test one week later.

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