Iiproteomics
Iimbonakalo
●Iiproteomics ezisemgangathweni: Isebenzisa i-LC-MS/MS ukuchonga ukwakheka kweproteni kwiisampuli ezintsonkothileyo (ii-SDS-PAGE gel strips, i-IP, i-Co-IP, i-Pull-down). Iingenelo: Akukho mda wenani lesampuli, ukufunyanwa ngokukhawuleza, ukucutshungulwa okulula kwesampuli, ukuveliswa okuphezulu, kunye nokukwazi ukubona iiproteni ezincinci.
● Iiproteomics zoBuninzi ezingenalebheli: Itekhnoloji engabhalwanga ilebheli enokufunyanwa kwesampuli nganye. Ilinganisa iiproteni ngokuthelekisa amandla esignali yepeptide kwidatha ye-mass spectrometry. Iingenelo: Ukusebenza okulula, ukuphumelela okuphezulu (akukho mda wenani lesampuli), kunye nokusetyenziswa okubanzi (kulungele ukuthelekiswa kweproteni "yobukho/ukungabikho" kwiintlobo ngeentlobo).
● I-DIA Quantitative Proteomics: Isebenzisa imo yokufunyanwa kwedatha (DIA), iskena zonke ii-ion kwiifestile ezihlukanisiweyo ukuze ibambe ulwazi olupheleleyo lwee-ion. Iingenelo: Ukuphindaphinda okungcono, ukugubungela okuphezulu kweproteni, kunye nokulinganisa okuchanekileyo kunemo ye-DDA (TMT/Free Label), efanelekileyo kwizifundo ezinkulu.
● I-4D-Label Free Quantitative Proteomics/4D-DIA Quantitative Proteomics: Isekelwe kwi-Bruker timsTOF mass spectrometer, yongeza ukuhamba kwe-ion (icandelo eliwela ungqubano) kulwahlulo lwe-3D lwendabuko. Iingenelo: Ukusetyenziswa okuphuculweyo kunye nokuchaneka kwe-ion, ukuphuculwa okubanzi kobunzulu bokugubungela, uvakalelo kunye nokufikelela; ubunzulu bokuchonga obuphezulu kunendlela ye-3D yendabuko.
● I-Astral Label Free Quantitative Proteomics/Astral DIA Quantitative Proteomics: Isekelwe kwi-OrbitrapTM AstralTM high-resolution mass spectrometer. Iingenelo: Ukuphuma okuphezulu (iisampuli ezili-100+/ngosuku olune-gradient yemizuzu esi-8), ukugubungela okunzulu (iiproteni ezingama-8,000+ ezifunyenwe kwiiseli zeHela kwimizuzu esi-8), kunye novakalelo oluphezulu (kufuneka isampuli encinci).
● Iiproteomics ze-TMT Quantitative: Isebenzisa iithegi ze-isotope ezili-18 ukulebhelisha iipeptides ukuze kufunyanwe iisampulu ezili-18 ngaxeshanye. Iingenelo: Uzinzo oluphezulu (ukuphazamiseka okuncinci kozinzo lwezixhobo, impazamo encinci yenkqubo) kunye novakalelo oluphezulu (ukwahlulwahlulwa kwezahlulo kunciphisa ubunzima besampulu, kuphucula ukufunyanwa kweproteni encinci).
● IiProteomics eziJongiweyo zePRM: Itekhnoloji ejolise kwisisombululo esiphezulu yokuchonga iiproteni/iipeptides ezijongiweyo. Iingenelo: Ivumela ukulinganiswa okulinganiselweyo/okupheleleyo kunye nokuqinisekiswa kweziphumo zeproteomics zobuninzi; akukho mingcelelo ye-antibody, ukusebenza ngokubanzi kuneWestern Blot kunye ne-ELISA.
Iingenelo
● Izixhobo Eziphambili: Zixhobise ngamaqonga eeproteomics aqhelekileyo kunye neespectrometers zobunzima obuphezulu (umz., i-4D, i-Astral).
● Ukuchonga Okuzinzileyo: Iinkqubo zokulawula umgangatho eziqinileyo ziqinisekisa iinkqubo zovavanyo ezihambelanayo nezizinzileyo.
● Iziphumo Ezithembekileyo: Uhlalutyo lomgangatho nolwamanani ngaxeshanye lubonelela ngenkcazo ehambelanayo, ubunzima beemolekyuli, ubuninzi kunye nolunye ulwazi oluphambili kwiqela ngalinye.
● Ukwenziwa kwezinto ngokuzenzekelayo okuphezulu: Iinkqubo ze-LC-MS ezizenzekelayo zivumela uhlalutyo olukhawulezayo kunye nokusebenza kakuhle kokwahlulahlula.
Iinkcukacha zeNkonzo
| Ukuthelekiswa kweeNkqubo zeProteomics ezingajoliswanga | ||||
| Ayinazo iileyibhile | I-DIA | I-TMT | ||
| Ukulebhelisha | NO | NO | EWE | |
| Imo Yokuskena Idatha | I-DDA | I-DDA | I-DIA | I-DDA |
| Ukuphinda kuvele | Ezantsi | Phezulu | Phezulu | |
| Uvelwano | Ezantsi | Phezulu | Phezulu | |
| Ukudibanisa izinto ezininzi | NO | EWE | NO | EWE |
| Isicelo | Ukuthelekisa ubukho/ukungabikho kweprotheyini | Iisampulu ezinkulu | Uhlalutyo lweebhetshi ezahlukeneyo zeesampuli | Ukuthelekisa ukubonakaliswa kweprotheni eyahlukileyo kuhlobo olufanayo kunye nezicubu |
| Ukuchaneka | ★ | ★★(4D/Astral DlA ★★★) | ★★ | |
| Inani lokuFunyanwa | ★ | ★★(4D/Astral D|A★★★) | ★★ | |
Iimfuno zeSampuli
Ngaba uyazibuza ukuba ingaba iisampulu zakho ziyahlangabezana na neekhrayitheriya zethu? Cofa apha ukuze ufumane ezethuiimfuno zamva nje zesampulu.
Ukuhamba komsebenzi wenkonzo
Ingqokelela yeeSampuli
Ukukhupha iMetabolite
Ukugaya ukutya nge-enzyme
Ukwahlukana
Ukufunyanwa kwedatha
Uhlalutyo lweDatha
Ukuhanjiswa Kwedatha
Iiproteomics zoMgangatho
1. Isisombululo seproteni/ijeli: itheyibhile yeziphumo ezisemgangathweni
2. Iziphumo zoKhangelo lweDatabase
2.1 Uluhlu lweziqendu zepeptide ezifaniswe kukhangelo lwedathabheyisi
2.2 Uluhlu lweeproteni ezifanisiweyo kuphendlo lwedathabheyisi
2.3 Uluhlu lweenguqu ezifaniswe nokukhangela kwisiseko sedatha (i-phosphorylation, i-ubiquitination, njl.njl.)
3. Idatha eluhlaza
Iiproteomics zobungakanani(Akukho ileyibhile/DIA/TMT)
1. Ukulungiswa kwangaphambili kwedatha
1.1 Ukukhangela kwiDatabase yeProteni
2. Uhlalutyo lokubonakaliswa kweeproteni
2.1 Uhlalutyo lweCandelo eliPhambili (i-PCA)
2.2 Ukuphambuka koMgangatho oQhelekileyo (RSD)
2.3 Ukusasazwa kweTrend ye-K-means
2.4 Uvavanyo lokuPhinda-phinda
2.5 Imephu yobushushu yokubonakalisa iiproteni
3. Isaziso esiSebenzayo
3.1 Isaziso esiSebenzayo se-GO
3.2 Isaziso esiSebenzayo se-KEGG
3.3 Isaziso esiSebenzayo se-COG
3.4 Isaziso esiSebenzayo seGOslim
3.5 Isaziso seDomain yoLwakhiwo lweProtheyini yePfam
4. Uhlalutyo lokwahluka kweeproteni (iikopi zebhayoloji ≥ 3)
4.1 Iziphumo zoHlalutyo lokwaHlukana kweeProteni
4.2 Ukusasazwa kotshintsho lwe-Differential Protein Fold (FC)
4.3 Uhlalutyo lwezibalo zoBuninzi beeProteni ezahlukeneyo
4.4 Iploti yeVolcano yeProteni eyahlukileyo
4.5 Imephu yobushushu yokudibanisa iiproteni ezahlukeneyo
4.6 Uhlalutyo lweProtheyini yoMntu oHlukileyo we-GO kunye noHlolo oluSebenzayo
4.7 Uhlalutyo lweProtheyini yokwahluka kwe-KEGG kunye noHlolo oluSebenzayo
4.8 Ingcaciso esebenzayo ye-COG yeProtheyini eyahlukileyo
4.9 Uhlalutyo lweProtheyini yeGOslim kunye noHlolo lweProtheyini eyahlukileyo
4.10 Uhlalutyo lweProtheyini yoLwahluko kunye noHlalutyo lweNdawo yokutyebisa
5. Uhlalutyo lokusebenzisana kweNethiwekhi yeeProteni
6. Ingcaciso yeProtein Reactome Pathway
7. Uqikelelo lweSignal Peptide
8. Ukufumaneka kweeProteni ezingaphantsi kweeseli
Iiproteomics zobungakanani(PRM)
1. Iziphumo zokulinganisa iiproteni ze-PRM
1.1 Isishwankathelo seziphumo zobungakanani
1.2 Ukusasazwa kweendawo ze-Fragment Ion Peak zePeptide Segments
Iiproteomics zobungakanani
1.Uhlalutyo lokubonakaliswa kweeproteni
↑Uhlalutyo lweeCandelo eziPhambili (i-PCA)↑Ukuphambuka okuMgangatho oQhelekileyo (RSD)
↑K-ithetha uSasazo lweNdlela ↑UlwalamanoAuhlalutyo lwePi-roteinEucinezeloLii-evels
↑ Imephu yobushushu bokubonakaliswa kweproteni
2. Isaziso esiSebenzayo
↑GO Isaziso esiSebenzayo↑Isaziso esiSebenzayo se-KEGG
↑ Isaziso esiSebenzayo se-COG ↑ Isaziso esiSebenzayo se-GOslim
↑ Isaziso seDomain yoBume beProtheyini yePfam
3.Uhlalutyo lokusebenzisana kwenethiwekhi yeeproteni
↑ Uhlalutyo lokusebenzisana kwenethiwekhi yeeproteni
4.Uqikelelo lweSignal Peptide
↑ Uqikelelo lweSignal Peptide
5. Ukufumaneka kweeProteni ezingaphantsi kweeseli
↑Ukubekwa kweeProteni kwiSeli
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