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No ke aha e lyse mua ai ka moʻomeheu cell i nā ʻulaʻula?

2022-12-23

Hoʻolauna kumu
ʻO ka Erythrocyte lysate kekahi o nā mea maʻalahi a maʻalahi e wehe i ke koko ʻulaʻula, ʻo ia hoʻi, ka hoʻokaʻawale ʻana i ke koko ʻulaʻula me ka lysate, ʻaʻole ia e hoʻopōʻino i nā cell nucleated a hiki ke hoʻopau piha i ke koko ʻulaʻula. ʻO ka Lysate cleavage kahi ʻano hoʻohemo ʻulaʻula ʻulaʻula maʻalahi, ka mea i hoʻohana nui ʻia no ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana o nā ʻiʻo ʻiʻo i hoʻopuehu ʻia e ka enzyme digestion, ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana o nā lymphocytes, a me ka wehe ʻana o nā ʻulaʻula ʻulaʻula i nā hoʻokolohua o ka protein tissue a me ka nucleic. ka ʻili ʻakika. ʻAʻole i loaʻa i nā ʻiʻo ʻiʻo i loaʻa i ka lysate o nā ʻulaʻula koko ʻulaʻula, a hiki ke hoʻohana hou ʻia no ka moʻomeheu mua, cell fusion, flow cytometry, kaʻawale a me ka unuhi ʻana o ka nucleic acid a me ka protein, etc.

Nā kuhikuhi no ka hoʻohana ʻana
Laʻana paʻi kiʻi
1. Ua hoʻokahe ʻia nā ʻiʻo hou e ka pancreas/enzyme a i ʻole collagenase a hoʻopuehu ʻia i loko o ka hoʻokuʻu ʻana o ke kelepona hoʻokahi, a ua hoʻolei ʻia ka supernatant e ka centrifugation.

2. E lawe i ka lysate ELS mai ka pahu hau ma 4â, e hoʻohui i ka lysate ELS i ka cell precipitate ma ka ratio o 1:3-5 (hoʻohui i 3-5ml o ka lysate i ka 1ml o ka cell compacted), e puhi mālie a hui pū.

3. Centrifuge ma 800-1000rpm no 5-8 mau minuke a hoʻolei i ka wai ʻulaʻula luna.

4. Ua hōʻiliʻili ʻia ka ʻaoʻao precipitated a centrifuged me ka Hank's solution a i ʻole serum-free culture solution no 2-3 mau manawa.

5, inā ʻaʻole paʻa ka paʻa ʻana, hiki ke hana hou i nā ʻanuʻu 2 a me 3.

6. Nā pūnaewele hoʻokuʻu hou no nā hoʻokolohua ma hope; Inā lawe ʻia ʻo RNA, ʻoi aku ka maikaʻi o ka hana ʻana pēlā ma ka hoʻonā i hoʻomākaukau ʻia mai ka ʻanuʻu 4 me ka wai DEPC

He pōkole loa ko ke koko ʻulaʻula, he 120 lā wale nō, akā, hoʻohua koke lākou i ke koko, a ma kēia ʻano, hiki iā lākou ke hoʻokaʻawale i nā cell, a ʻo lākou ka wikiwiki o ka puʻunaue o nā mea a pau, no laila he waiwai nui kēia cell. no laila he mea pono loa ia no ka moʻomeheu cell. He mea maʻalahi loa, ʻaʻohe ona organelles i loko, ʻo nā membrane cell wale nō a me nā protein.

Mamua:He aha nā hana o ka pahu ELISA?

Aʻe:Nā koi reagent a me ka hōʻike kumu o ka hoʻonā hoʻonā.