I-Electrophoresis: iyini, yenzelwe ini nokuthi yenziwa kanjani

-Delile

• Yenzelwe ini
• Kwenziwa kanjani
• Izinhlobo ze-electrophoresis
• 1. I-hemoglobin electrophoresis
• 2. Amaphrotheni electrophoresis

I-Electrophoresis inqubo yaselebhu eyenziwe ngenhloso yokuhlukanisa ama-molecule ngokobungako bawo kanye necala lokukhokhisa ugesi ukuze kutholakale ukuthi kutholakala izifo, ukuvezwa kwamaprotheni kungaqinisekiswa noma kungabonakala amagciwane amancane.

I-Electrophoresis inqubo elula nebiza kancane, esetshenziswa emisebenzini yelabhorethri nasemiklamweni yocwaningo. Ngokwenhloso ye-electrophoresis, kungadingeka ukwenza ezinye izivivinyo kanye nezivivinyo ukuze kufinyelelwe ukuxilongwa, isibonelo.

Yenzelwe ini

I-Electrophoresis ingenziwa ngezinhloso eziningana, zombili kumaphrojekthi ocwaningo nasekuxilongweni, ngoba kuyindlela elula nebiza kancane.Ngakho-ke, i-electrophoresis ingenziwa ku:

• Thola amagciwane, isikhunta, amabhaktheriya kanye nezimuncagazi, ngalolu hlelo lokusebenza luvame kakhulu kumaphrojekthi ocwaningo;
• Ukuhlolwa kobaba;
• Bheka ukubonakaliswa kwamaprotheni;
• Khomba ukuguquka kwezakhi zofuzo, ube wusizo ekuhlolweni kwama-leukemias, isibonelo;
• Hlaziya izinhlobo ze-hemoglobin ejikelezayo, ube wusizo ekutholeni i-sickle cell anemia;
• Hlola inani lamaprotheni akhona egazini.

Ngokwenhloso ye-electrophoresis, kungadingeka ukwenza ezinye izivivinyo ezihambisanayo kudokotela ukuqedela ukuxilongwa.

Kwenziwa kanjani

Ukwenza i-electrophoresis kuyadingeka i-gel, engaba nge-polyacrylamide noma i-agarose ngokuya ngenhloso, i-electrophoresis buffer ne-vat, umaka wesisindo samangqamuzana nodayi we-fluorescent, ngaphezu kwe-UV noma imishini yokukhanyisa ye-LED, eyaziwa nangokuthi i-transilluminator .

Ngemuva kokulungisa ijeli, kufanele kubekwe into ethize yokwenza imithombo yejel, edume ngokubizwa ngokuthi ikama, bese ibeka ijeli. Lapho ijeli selilungile, vele ufake izinto emithonjeni. Ngalokhu, umaka wesisindo samangqamuzana kufanele abekwe komunye wemithombo, ukulawula okuhle, okuyisithako esaziwa ukuthi siyini, isilawuli esingesihle, esiqinisekisa ukusebenza kwempendulo, namasampuli azohlaziywa. Onke amasampula kufanele ahlanganiswe nedayi ye-fluorescent, ngoba ngale ndlela kungenzeka ukubona ngeso lengqondo amabhande ku-transilluminator.

Ijeli enamasampula kufanele ibekwe emgodini we-electrophoresis, oqukethe isixazululo esithile se-buffer, bese idivayisi ivulwa ukuze kube namandla kagesi futhi, ngenxa yalokho, umehluko ongaba khona, obalulekile ekwehlukaneni kwezinhlayiya ngokusho kumthwalo wazo nosayizi. Isikhathi sokusebenza se-electrophoretic siyahluka ngokwenqubo yenqubo, futhi singahlala kufika ehoreni elilodwa.

Ngemuva kwesikhathi esinqunyiwe, kungenzeka ukubuka umphumela we-electrophoretic run ngokusebenzisa i-transilluminator. Lapho ijeli lifakwa ngaphansi kwelambu le-UV noma le-LED, kuyenzeka ubone iphethini yokubopha: lapho i-molecule iba nkulu, kuncane lapho ifuduka, isondela emthonjeni, ngenkathi i-molecule ikhanya, amandla amakhulu okufuduka ayanda.

Ukuze impendulo iqinisekiswe, kuyadingeka ukuthi amabhande we-positive control aboniswe nokuthi ekulawuleni okungalungile akukho okubonwayo, ngoba uma kungenjalo kuyinkomba yokuthi bekukhona ukungcola, futhi yonke inqubo kufanele iphindwe.

Izinhlobo ze-electrophoresis

I-Electrophoresis ingenziwa ngezinhloso ezahlukahlukene futhi, ngokwenhloso yayo, izinhlobo eziningi zejel zingasetshenziswa, okuvame kakhulu ukuba yi-polyacrylamide ne-agarose.

I-Electrophoresis ukukhomba ama-microorganism kuvame ukwenziwa kumalabhorethri ocwaningo, kepha, ngezinhloso zokuxilonga, i-electrophoresis ingasetshenziselwa ukukhomba izifo nezifo ze-hematological eziguquguquka ngokwanda kwenani lamaprotheni, okuyizinhlobo eziyinhloko ze-electrophoresis:

1. I-hemoglobin electrophoresis

I-Hemoglobin electrophoresis inqubo yaselebhu eyenzelwa ukukhomba izinhlobo ezahlukahlukene ze-hemoglobin ejikeleza egazini, okwenza kube lula ukubona ubukhona bezifo ezihlobene nokuhlanganiswa kwe-hemoglobin. Uhlobo lwe-hemoglobin lukhonjwa ngokusebenzisa i-electrophoresis ku-pH ethile, kahle phakathi kuka-8.0 no-9.0, ngephethini yamabhande eqinisekisiwe angaqhathaniswa nephethini ejwayelekile, evumela ukutholakala kobukhona bama-hemoglobin angajwayelekile.

Yenzelwe ini: I-hemoglobin electrophoresis yenziwa ukuphenya nokuhlola izifo ezihlobene ne-hemoglobin synthesis, njenge-sickle cell anemia kanye nesifo se-hemoglobin C, ngaphezu kokuba lusizo ekuhlukaniseni i-thalassemia. Funda indlela yokuhumusha i-hemoglobin electrophoresis.

2. Amaphrotheni electrophoresis

Amaphrotheni electrophoresis ukuhlolwa okucelwe ngudokotela ukuhlola inani lamaprotheni ajikeleza egazini, ngaleyo ndlela, ukukhomba izifo. Lokhu kuhlolwa kwenziwa kusampula yegazi, ekhishwe ngamasenthimitha ukuthola i-plasma, okuyingxenye yegazi, equkethe, phakathi kwezinye izinto, zamaprotheni.

Ngemuva kwe-electrophoresis, iphethini yamabhande angabonwa futhi, ngemuva kwalokho, igrafu lapho kukhonjiswa khona ubuningi beqhezu ngalinye lamaprotheni, okuyisisekelo sokuxilongwa.

Yenzelwe ini: Amaphrotheni electrophoresis avumela udokotela ukuthi aphenye ngokuvela kwe-myeloma amaningi, ukuphelelwa amandla kwamanzi, ukuqina kwesibindi, ukuvuvukala, isifo sesibindi, i-pancreatitis, i-lupus kanye nomfutho wegazi ophezulu ngokusho kwephethini yebhande negrafu elethwe embikweni wokuhlolwa.

Qonda ukuthi kwenziwa kanjani nokuthi ungayiqonda kanjani imiphumela ye-protein electrophoresis.