ELISA Cov Khoom Siv ua haujlwm li cas

Ob chav antibody sandwich txoj kev 1. Pob: Nrog 0.05MPH9. cov carbonate pob yog buffered kom dilute cov tshuaj tiv thaiv rau cov ntsiab lus protein ntawm 1 mus rau 10 ?g / ml. Ntxiv 0.1 ml, 4 degrees C thaum hmo ntuj, nyob rau hauv cov tshuaj tiv thaiv qhov ntawm txhua lub phaj polystyrene. Hnub tom qab, muab cov tshuaj pov tseg rau hauv lub qhov thiab ntxuav nws 3 zaug nrog kev ntxhua khaub ncaws rau 3 feeb txhua zaus.

(rau luv luv, ntxuav, tib yam hauv qab no). 2. Ntxiv cov qauv: Ntxiv ib qho kev dilution ntawm cov qauv mus kuaj 0.1 ml nyob rau hauv cov saum toj no - hais txog ntim cov tshuaj tiv thaiv qhov, incubate rau 1 teev ntawm 37 degrees C. Tom qab ntawd ntxuav.

(Tam sim no ua qhov khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob khoob

Incubate 0.5 mus rau 1 teev ntawm 37 degrees C thiab ntxuav.

4. Ntxiv substrate kua xim zaub: ntxiv ib ntus kev npaj ntawm TMB substrate tov 0.1 ml, 37 degrees C 10 mus rau 30 feeb nyob rau hauv txhua lub qhov tshuaj tiv thaiv.

5. Cov tshuaj tiv thaiv kawg: Ntxiv 2M sulfuric acid 0.05 ml rau txhua qhov tshuaj tiv thaiv . 6. Cov txiaj ntsig tau txiav txim siab: tuaj yeem ua rau ntawm lub ntsej muag dawb, ncaj qha nrog lub qhov muag liab qab los soj ntsuam cov txiaj ntsig: cov xim tsaus dua hauv qhov qhov tshuaj tiv thaiv, qhov muaj zog dua qhov zoo, cov tshuaj tiv thaiv tsis zoo yog xim tsis muaj xim los yog ntiav heev, raws li cov xim yog reflected nyob rau hauv qhov tob, rau cov xim ntawm qhov tob.

Od tus nqi scanted: ntawm ELISA tester, ntawm 450nm (yog tias BTS xim rendering, ces 410nm), kom dawb paug tswj qhov xoom ingested los ntsuas tus nqi OD ntawm txhua lub qhov, yog ntau tshaj qhov teev tsis zoo tswj OD tus nqi ntawm 2.1 npaug, uas yog, zoo.

Txoj cai tsis ncaj

1. Siv lub pob yuav tsum tau buffered kom dilute cov paub antigen rau 1 mus rau 10 mg / ml, ib lub qhov ntxiv 0.1 ml, 4 degrees C thaum hmo ntuj;

2. Ntxuav 3 zaug rau hnub tom qab;

3. Ntxiv ib qho kev dilution ntawm cov qauv (tsis paub cov tshuaj tiv thaiv) 0.1 ml nyob rau saum toj no - hais txog lub qhov ntim cov tshuaj tiv thaiv, incubate rau 1 teev ntawm 37 degrees C, ntxuav;

4. (ob qho tib si dawb paug, tsis zoo thiab qhov zoo tswj lub qhov) nyob rau hauv cov tshuaj tiv thaiv qhov, ntxiv ib tug freshly diluted enzyme daim ntawv lo thib ob antibody (anti-antibody) 0.1 ml;

5.37 ° C incubation 35-60 feeb, ntxuav;

6.'Lub sijhawm kawg koj ntxuav nrog DDW. Cov kauj ruam ntxiv yog tib yam li "ob chav antibody sandwich txoj kev" ntawm 4, 5, 6.