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Vim li cas cell kab lis kev cai lyse qe ntshav liab ua ntej?

2022-12-23

Kev taw qhia yooj yim
Erythrocyte lysate yog ib txoj hauv kev yooj yim tshaj plaws thiab yooj yim tshem tawm cov qe ntshav liab, uas yog, faib cov qe ntshav liab nrog lysate, uas tsis ua rau cov hlwb nucleated thiab tshem tawm cov qe ntshav liab tag nrho. Lysate cleavage yog ib txoj kev tshem tawm cov ntshav liab me me, uas feem ntau yog siv rau kev sib cais thiab ua kom huv ntawm cov ntaub so ntswg dispersed los ntawm kev zom cov enzyme, kev sib cais thiab purification ntawm lymphocytes, thiab tshem tawm cov qe ntshav liab hauv kev sim ntawm cov nqaijrog protein thiab nucleic. kua qaub extraction. Cov ntaub so ntswg tau txais los ntawm lysate ntawm cov qe ntshav liab tsis muaj cov qe ntshav liab, thiab tuaj yeem siv ntxiv rau kev coj noj coj ua, cell fusion, ntws cytometry, sib cais thiab rho tawm cov nucleic acid thiab protein, thiab lwm yam.

Cov lus qhia rau kev siv
Cov ntaub so ntswg cell qauv
1. Cov ntaub so ntswg tshiab tau zom los ntawm pancreas / enzyme los yog collagenase thiab dispersed rau hauv ib leeg cell suspension, thiab cov supernatant raug muab pov tseg los ntawm centrifugation.

2. Nqa ELS lysate los ntawm lub tub yees ntawm 4â ¢, ntxiv ELS lysate rau cell precipitate hauv qhov piv ntawm 1: 3-5 (ntxiv 3-5ml ntawm lysate rau 1ml ntawm cell compacted), maj mam tshuab thiab sib tov.

3. Centrifuge ntawm 800-1000rpm rau 5-8 feeb thiab muab pov tseg cov kua liab liab ntshiab.

4. Lub precipitated ib feem tau sau thiab centrifuged nrog Hank cov tshuaj los yog cov kab lis kev cai tsis muaj ntshav qab zib rau 2-3 zaug.

5, yog tias qhov tawg tsis tiav / ua tiav tuaj yeem rov ua cov kauj ruam 2 thiab 3.

6. Cov hlwb rov ua haujlwm rau kev sim tom ntej; Yog tias RNA raug rho tawm, nws yog qhov zoo tshaj rau ua li ntawd hauv cov tshuaj npaj los ntawm Kauj Ruam 4 siv dej DEPC

Cov qe ntshav liab muaj lub neej luv luv, tsuas yog 120 hnub xwb, tab sis lawv rov tsim cov ntshav sai heev, thiab qhov no lawv muaj peev xwm tshwj xeeb ntawm cov cell faib, thiab lawv yog cov faib cov hlwb ceev tshaj plaws ntawm txhua tus, yog li lub xov tooj ntawm tes no tseem ceeb heev, yog li nws tseem ceeb heev rau kab lis kev cai ntawm tes. Nws yooj yim heev, nws tsis muaj organelles nyob rau hauv nws, tsuas yog cell membranes thiab proteins.

Yav dhau los:ELISA cov khoom siv yog dab tsi?

Tom ntej no:Reagent yuav tsum tau thiab cov sawv cev tseem ceeb ntawm cov tshuaj concentration.