Ama-Proteomics
Izici
●Ama-Proteomics Asezingeni Elifanele: Isebenzisa i-LC-MS/MS ukuhlonza ukwakheka kwamaprotheni kumasampula ayinkimbinkimbi (ama-SDS-PAGE gel strips, i-IP, i-Co-IP, i-Pull-down). Izinzuzo: Akukho mkhawulo wenombolo yesampula, ukutholwa okusheshayo, ukucubungula okulula kwesampula, ukukhishwa okuphezulu, kanye nekhono lokuthola amaprotheni amaningi kakhulu.
● Ama-Proteomics Okulinganisa Angenamalebula: Ubuchwepheshe obungenamalebula obunokutholwa kwesampula ngayinye. Ilinganisa amaprotheni ngokuqhathanisa amandla esignali ye-peptide kudatha ye-mass spectrometry. Izinzuzo: Ukusebenza okulula, ukuphuma okuphezulu (akukho mkhawulo wenombolo yesampula), kanye nokusebenza okubanzi (kufanelekela ukuqhathaniswa kweprotheni yokuhlukanisa "yokuba khona/ukungabikho" kuzo zonke izinhlobo).
● I-DIA Quantitative Proteomics: Isebenzisa imodi yokuthola idatha (DIA) engaxhomekile kuyo, iskena wonke ama-ion kumafasitela ahlukaniswe ukuze ibambe ulwazi oluphelele lwama-ion. Izinzuzo: Ukuphindaphinda okungcono, ukumbozwa okuphezulu kwamaprotheni, kanye nokulinganisa okunembe kakhulu kunemodi ye-DDA (i-TMT/engenayo ilebula), efanelekile ezifundweni ezinkulu.
● I-4D-Label Free Quantitative Proteomics/4D-DIA Quantitative Proteomics: Isekelwe ku-Bruker timsTOF mass spectrometer, enezela ukuhamba kwe-ion (isigaba sokushayisana) ekuhlukanisweni kwendabuko kwe-3D. Izinzuzo: Ukusetshenziswa okuthuthukisiwe kanye nokunemba kwe-ion, ukuthuthukiswa okuphelele ekujuleni kokumbozwa, ukuzwela kanye nokusetshenziswa; ukujula kokuhlonza okuphezulu kunendlela yendabuko ye-3D.
● I-Astral Label Free Quantitative Proteomics/Astral DIA Quantitative Proteomics: Isekelwe ku-OrbitrapTM AstralTM high-resolution mass spectrometer. Izinzuzo: Ukuphuma okuphezulu (amasampula ayi-100+/ngosuku nge-gradient yemizuzu engu-8), ukumbozwa okujulile (amaprotheni angu-8,000+ atholakale kumaseli e-Hela ngemizuzu engu-8), kanye nokuzwela okuphezulu (isampula encane iyadingeka).
● I-TMT Quantitative Proteomics: Isebenzisa amathegi e-isotope angu-18 ukulebula ama-peptide ukuze kutholakale amasampula angu-18 ngesikhathi esisodwa. Izinzuzo: Ukuqina okuphezulu (ukuphazamiseka okuncane kokuqina kwezinsimbi, iphutha elincane lesistimu) kanye nokuzwela okuphezulu (ukuhlukanisa izingxenye kunciphisa ubunzima besampula, kuthuthukisa ukutholwa kwamaprotheni amaningi).
● Ama-Proteomics Ahlosiwe e-PRM: Ubuchwepheshe obuhlosiwe obunesinqumo esiphezulu sokuthola amaprotheni/ama-peptide ahlosiwe ngokukhetha. Izinzuzo: Ivumela ukulinganiswa okuhlobene/okuphelele kanye nokuqinisekiswa kwemiphumela ye-proteomics yobuningi; akukho ukuvinjelwa kwama-antibody, ukusebenza okubanzi kune-Western Blot kanye ne-ELISA.
Izinzuzo
● Imishini Ethuthukisiwe: Hlomisa ngamapulatifomu avamile e-proteomics kanye nama-spectrometer esisindo athuthukisiwe (isib. i-4D, i-Astral).
● Ukutholwa Okuzinzile: Izinhlelo zokulawula ikhwalithi eqinile ziqinisekisa izinqubo zokuhlola ezihambisanayo nezizinzile.
● Imiphumela Ethembekile: Ukuhlaziywa kwekhwalithi kanye nokwezibalo ngesikhathi esisodwa kunikeza ukubonakaliswa okuhlobene, isisindo sama-molecule, ubuningi kanye nolunye ulwazi olubalulekile lweqembu ngalinye.
● Ukuzenzakalela Okuphezulu: Izinhlelo ze-LC-MS ezizenzakalelayo zivumela ukuhlaziywa okusheshayo kanye nokusebenza okuhle kakhulu kokuhlukaniswa.
Imininingwane Yesevisi
| Ukuqhathaniswa Kwamasu E-Proteomics Angahlosiwe | ||||
| Ayinawo amalebula | I-DIA | I-TMT | ||
| Ukulebula | NO | NO | YEBO | |
| Imodi Yokuskena Idatha | I-DDA | I-DDA | I-DIA | I-DDA |
| Ukuphinda kukhiqizwe | Ngaphansi | Phezulu | Phezulu | |
| Ukuzwela | Ngaphansi | Phezulu | Phezulu | |
| Ukuphindaphinda | NO | YEBO | NO | YEBO |
| Isicelo | Ukuqhathanisa ukuba khona/ukungabikho kwamaprotheni | Amasampula amakhulu | Ukuhlaziywa kwamaqoqo ahlukene amasampula | Ukuqhathanisa ukubonakaliswa kweprotheni okuhlukile ohlotsheni olufanayo kanye nezicubu |
| Ukunemba | ★ | ★★(4D/Astral DlA ★★★) | ★★ | |
| Inani Lokutholwa | ★ | ★★(4D/Astral D|A★★★) | ★★ | |
Izidingo Zesampula
Uyazibuza ukuthi amasampula akho ayahlangabezana yini nezindinganiso zethu? Chofoza lapha ukuze uthole ezethuizidingo zakamuva zesampula.
Ukugeleza Komsebenzi Wesevisi
Iqoqo Lesampula
Ukukhishwa Kwamaprotheni
Ukugaya ukudla nge-enzyme
Ukuhlukana
Ukutholwa Kwedatha
Ukuhlaziywa Kwedatha
Ukulethwa Kwedatha
Ama-Proteomics Ekhwalithi
1. Isixazululo/ijeli yephrotheni: ithebula lemiphumela esezingeni eliphezulu
2. Imiphumela Yokusesha Yesizindalwazi
2.1 Uhlu lwezingxenye ze-peptide ezifaniswe ekusesheni kwedatha
2.2 Uhlu lwamaprotheni afaniswe ekusesheni kwedatha
2.3 Uhlu lwezinguquko ezifaniswe ekusesheni kwedatha (i-phosphorylation, i-ubiquitination, njll.)
3. Idatha eluhlaza
Ama-Proteomics Obuningi(Akukho Lebula/DIA/TMT)
1. Ukucubungula Idatha Ngaphambi Kokuqala
1.1 Usesho Lwesizindalwazi Samaprotheni
2. Ukuhlaziywa Kokubonakaliswa Kwamaprotheni
2.1 Ukuhlaziywa Kwezingxenye Eziyinhloko (i-PCA)
2.2 Ukuphambuka Okujwayelekile Okuhlobene (RSD)
2.3 Ukusatshalaliswa Kwethrendi Okungu-K
2.4 Ukuhlolwa Kokuphindeka Kabusha
2.5 Imephu Yokushisa Yokubonakaliswa Kwamaprotheni
3. Isichasiselo Esisebenzayo
3.1 Isaziso Esisebenzayo se-GO
3.2 Isichasiselo Sokusebenza se-KEGG
3.3 Isichasiselo Sokusebenza se-COG
3.4 Isichasiselo Esisebenzayo se-GOslim
3.5 Isakhiwo sePfam Protein Isichasiselo Sesizinda
4. Ukuhlaziywa Kokwehluka Kwamaprotheni (okuphindaphindwayo kwezinto eziphilayo ≥ 3)
4.1 Imiphumela Yokuhlaziya Ukwehluka Kwamaprotheni
4.2 Ukusatshalaliswa Kwezinguquko Ezihlukahlukene Ze-Protein Fold (FC)
4.3 Ukuhlaziywa Kwezibalo Zobuningi Bamaphrotheni Ahlukile
4.4 Isiqephu Sentaba-mlilo Esihlukile Seprotheyini
4.5 Imephu Yokushisa Ehlukanisayo Yamaphrotheni Ahlukene
4.6 Ukuhlaziywa Kwephrotheni Ehlukile Ye-GO Functional Annotation kanye Nokucebisa
4.7 Ukuhlaziywa Kwephrotheni Ehlukile ye-KEGG kanye Nokucebisa
4.8 Isichasiselo Sokusebenza Se-COG Yeprotheyini Ehlukile
4.9 Ukuhlaziywa Kwe-Differential Protein GOslim Annotation kanye Nokucebisa
4.10 Ukuhlaziywa Kwesakhiwo Sephrotheni Esihlukile Nokucebisa Isizinda
5. Ukuhlaziywa Kokuxhumana Kwamaphrotheni Nenethiwekhi
6. Isichasiselo se-Protein Reactome Pathway
7. Ukubikezela kwe-Signal Peptide
8. Ukutholakala Kwamaphrotheni Angaphansi Kwamaseli
Ama-Proteomics Obuningi(I-PRM)
1. Imiphumela Yokulinganisa Amaprotheni e-PRM
1.1 Ukubuka Konke Kwemiphumela Yokulinganisa
1.2 Ukusatshalaliswa Kwezindawo Eziphakeme Ze-Fragment Ion Zezingxenye Ze-Peptide
Ama-Proteomics Obuningi
1.Ukuhlaziywa Kokubonakaliswa Kwamaprotheni
↑Ukuhlaziywa Kwezingxenye Eziyinhloko (i-PCA)↑Ukuphambuka Okujwayelekile Okuhlobene (RSD)
↑K-isho Ukusatshalaliswa Kwethrendi ↑UkuhambisanaAukuhlolwa kwePi-roteinEukucindezelaLama-evel
↑ Imephu Yokushisa Yokubonakaliswa Kwamaprotheni
2. Isichasiselo Esisebenzayo
↑GO Isichasiselo Esisebenzayo↑Isichasiselo Sokusebenza se-KEGG
↑ Isichasiselo Sokusebenza se-COG ↑ Isichasiselo Sokusebenza se-GOslim
↑ Isichasiselo Sesizinda Sesakhiwo Sephrotheni ye-Pfam
3.Ukuhlaziywa Kokuxhumana Kwamaphrotheni Nenethiwekhi
↑ Ukuhlaziywa Kokusebenzisana Kwenethiwekhi Yamaprotheni
4.Ukubikezela kwe-Signal Peptide
↑ Ukubikezela kwe-Signal Peptide
5. Ukutholakala Kwamaphrotheni Angaphansi Kwamaseli
↑Ukwakheka Kwamaprotheni Angaphansi Kwamaseli
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