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Kedu ihe kpatara omenala cell ji ebu ụzọ lyse mkpụrụ ndụ ọbara uhie?

2022-12-23

Okwu mmalite
Erythrocyte lysate bụ otu n'ime ụzọ kachasị mfe ma dị mfe isi wepụ mkpụrụ ndụ ọbara uhie, ya bụ, iji lysate kewaa mkpụrụ ndụ ọbara uhie, nke na-adịghị emebi mkpụrụ ndụ ndị nwere oghere ma nwee ike iwepụ mkpụrụ ndụ ọbara uhie kpamkpam. Lysate cleavage bụ usoro mwepụ mkpụrụ ndụ ọbara uhie dị nro, nke a na-ejikarị maka nkewa na ịdị ọcha nke mkpụrụ ndụ anụ ahụ gbasasịrị site na mgbaze enzyme mgbaze, nkewa na ịdị ọcha nke lymphocytes, na mwepụ nke mkpụrụ ndụ ọbara uhie na nnwale nke protein anụ ahụ na nucleic. mmịpụta acid. Mkpụrụ ndụ anụ ahụ enwetara site na lysate nke mkpụrụ ndụ ọbara uhie enweghị mkpụrụ ndụ ọbara uhie, a pụkwara iji ya mee ihe maka ọdịbendị mbụ, njikọ cell, cytometry na-asọpụta, nkewa na mmịpụta nke nucleic acid na protein, wdg.

Ntuziaka maka ojiji
Ihe atụ cell cell
1. A na-agbari anụ ahụ ọhụrụ site na pancreas / enzyme ma ọ bụ collagenase wee gbasasịa n'ime otu nkwusioru cell, na centrifugation tụfuru ihe dị elu.

2. Were ELS lysate si na refrjiraeto na 4â, tinye ELS lysate na cell precipitate na nha nke 1: 3-5 (tinye 3-5ml nke lysate na 1ml nke cell compacted), jiri nwayọọ fesaa na mix.

3. Centrifuge na 800-1000rpm maka nkeji 5-8 ma tụfuo mmiri mmiri dị ọcha na-acha uhie uhie.

4. A nakọtara akụkụ ahụ ewepụtara ma were ihe ngwọta Hank ma ọ bụ ihe ngwọta omenala na-enweghị serum maka oge 2-3.

5, ma ọ bụrụ na mgbawa ezughị oke / zuru ezu, enwere ike ịmegharị usoro 2 na 3 ọzọ.

6. Mkpụrụ ndụ mweghachi maka nyocha ndị na-esote; Ọ bụrụ na ewepụtara RNA, ọ kacha mma ime ya na ngwọta a kwadebere site na Nzọụkwụ 4 iji mmiri DEPC

Mkpụrụ ndụ ọbara uhie nwere okirikiri ndụ dị mkpụmkpụ, naanị ụbọchị 120, mana ha na-amụba ọbara ngwa ngwa, na nke a, ha nwere ike nkewa nkewa, na ha bụ sel na-ekesa ngwa ngwa, yabụ sel a bara ezigbo uru. ya mere ọ bara uru nke ukwuu maka omenala cell. Ọ dị nnọọ mfe, ọ nweghị akụkụ ahụ dị n'ime ya, naanị membranes cell na protein.

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