Kukho amaqhawe amaninzi angadunyiswanga asingqongileyo, abonakala ngathi aqhelekile, kodwa eneneni anegalelo elikhulu kuthi ngokuthe cwaka. I-Proteinase K "liqhawe elingadunyiswanga" kwishishini lokuxilongwa kweemolekyuli, nangona xa ithelekiswa "nelikhulu nelinamandla" kwishishini, i-proteinase K iphantsi kakhulu kangangokuba kudala singayinaki ukubaluleka kwayo. Ngokuqhambuka kobhubhane omtsha we-crown, imfuno ye-proteinase K inyukile, kwaye ukubonelelwa ekhaya nakwamanye amazwe kusemva kakhulu kokusetyenziswa, kwaye wonke umntu ngequbuliso waqonda ukuba i-proteinase K ibaluleke kakhulu.
Yintoni ukusetyenziswa kweproteinase K?
I-Proteinase K yi-serine protease enomsebenzi we-proteolytic enzyme kwaye inokugcina umsebenzi kwiindawo ezahlukeneyo (i-pH (4-12.5), i-buffer enetyuwa eninzi, ubushushu obuphezulu be-70°C, njl.njl.). Ukongeza, umsebenzi we-proteinase K awuthintelwa yi-SDS, i-urea, i-EDTA, i-guanidine hydrochloride, i-guanidine isothiocyanate, njl.njl., kwaye isixa esithile sesepha sinokunyusa umsebenzi we-proteinase K. Kunyango lwezonyango (intsholongwane kunye nokubulala iintsholongwane), ukutya (ukuthambisa inyama), ulusu (ukuthambisa iinwele), ukwenza iwayini (ukucaciswa kotywala), ukulungiswa kwe-amino acid (iintsiba eziwohlokileyo), ukukhutshwa kwe-nucleic acid, i-in situ hybridization, njl.njl., i-proteinase K Kukho izicelo. Ukusetyenziswa okuqhelekileyo kukukhutshwa kwe-nucleic acid.
I-Proteinase K ingenza i-enzyme zonke iintlobo zeeproteni kwisampuli, kuquka ezo histones zibotshelelwe ngokuqinileyo kwi-nucleic acids, ukuze ii-nucleic acids zikhutshwe kwisampuli zize zikhutshwe kwisicatshulwa, nto leyo eyenza kube lula inyathelo elilandelayo lokukhupha nokucoca. Ekufumaneni i-viral nucleic acid, i-proteinase K yenye yezona zinto zibalulekileyo kwisisombululo sokufumana isampuli yentsholongwane. I-Proteinase K inokuqhekeka kwaye ingasebenzi i-coat protein yentsholongwane, nto leyo ikhuselekileyo ngexesha lokuthutha nokufumanisa; ukongeza, i-proteinase K inokonakalisa i-RNase. Ithintela ukonakala kwe-viral RNA kwaye iququzelele ukufunyanwa kwe-nucleic acid.
Udumo olukhawulezileyo lweproteinase K
Nokuba kusentsimini yophando lwesayensi okanye kwicandelo le-IVD, ukukhupha i-nucleic acid yeyona nto ibalulekileyo, ngoko ke i-proteinase K ibisoloko iyinto ebalulekileyo. Nangona kunjalo, ngaphambili, i-proteinase K yayingaziwa kangako kunendima yayo. Inxalenye enkulu yoku yayikukuba ubudlelwane bokubonelela kunye nemfuno ye-proteinase K babuzinzile kakhulu. Bambalwa abantu abanokucinga ukuba ukubonelela nge-proteinase K kuya kuba yingxaki.
Ngokuqhambuka kobhubhane omtsha we-crown, imfuno yovavanyo lwe-nucleic acid inyukile. Ukusukela ekupheleni kukaJuni ka-2020, iTshayina igqibe uvavanyo olutsha lwe-crown oluphantse lube zizigidi ezingama-90, kwaye eli nani liyoyikisa nangakumbi kwihlabathi liphela. Kwiimvavanyo zokukhupha i-nucleic acid, uxinzelelo olusebenzayo lwe-proteinase K lumalunga ne-50-200 μg/mL. Ngokubanzi, kuthatha malunga ne-100 μg ye-proteinase K ukukhupha isampuli ye-nucleic acid. Ekusetyenzisweni kwangempela, ukuze kwandiswe ukusebenza kakuhle kokukhupha i-nucleic acid, rhoqo i-Proteinase K iya kusetyenziswa ngobuninzi. Ukufunyanwa kwe-nucleic acid ye-coronavirus entsha kuzise inani elikhulu lemfuno ye-proteinase K. Umlinganiselo wokuqala wokubonelela kunye nemfuno ye-proteinase K waphulwa ngokukhawuleza, kwaye i-proteinase K yaba yinto ebalulekileyo yokuthintela ubhubhane ngobusuku obunye.
Ubunzima ekuvelisweni kweproteinase K
Nangona ngokukhula kobhubhane, ixabiso elibalulekileyo leproteinase K liye laxatyiswa ngabantu, kuyadanisa ukuba ngenxa yokuba iproteinase K incinci kakhulu, zimbalwa iinkampani zasekhaya ezibandakanyekileyo ekuvelisweni kweproteinase K. Xa abantu befuna ukuseka imveliso yeproteinase K Ngexesha lenkqubo yemveliso, kwafunyaniswa ukuba iproteinase K yiprotein ekhethekileyo kakhulu. Kunzima kakhulu ukwandisa amandla emveliso yeproteinase K ngexesha elifutshane.
Ukuveliswa okukhulu kweproteinase K kujongene nobunzima obulandelayo
1. Ukubonakaliswa okuphantsi
I-Proteinase K inokonakalisa iiproteni ezininzi ngaphandle kokuphazamisa ngokukodwa kwaye ibangele ubuthi obukhulu kwiseli yomzi wokubonakaliswa. Ke ngoko, inqanaba lokubonakaliswa kwe-proteinase K ngokubanzi liphantsi kakhulu. Ukuhlolwa kweenkqubo zokubonakaliswa kunye neentlobo ze-proteinase K eziveliswa kakhulu ngokubanzi kufuna umjikelo omde.
2. Iintsalela zeepigment kunye neenucleic acids
Ukubiliswa okukhulu kungenisa inani elikhulu leentsalela ze-pigment kunye ne-nucleic acid. Kunzima ukususa ezi zinto zingcolileyo ngenkqubo elula yokucoca, kwaye ukucocwa okuntsonkothileyo kwandisa iindleko kwaye kunciphisa izinga lokubuyiselwa.
3. Ukungazinzi
I-Proteinase K ayizinzanga ngokwaneleyo, ingazincipha ngokwayo, kwaye kunzima ukuyigcina ngokuzinzileyo kwi-37°C ixesha elide ngaphandle kwesixhobo sokukhusela.
4. Kulula ukukhawulezisa
Xa ulungiselela umgubo we-proteinase K omisiweyo ngokuqandisa, ukuqinisekisa ukuba umxholo oqinileyo we-proteinase K kumgubo owomisiweyo ngokuqandisa mkhulu, kuyimfuneko ukongeza i-arhente yokukhusela eyomisiweyo ngokuqandisa kuxinzelelo oluphezulu, kodwa xa uxinzelelo lwe-proteinase K lufikelela kwi-20mg/mL nangaphezulu, kulula Ukuhlanganiswa kwenza i-precipitate, nto leyo ezisa ubunzima obukhulu ekuqandiseni ukomisa kwe-proteinase K ngomxholo ophezulu oqinileyo.
5. Utyalo-mali olukhulu
I-Proteinase K inomsebenzi omkhulu we-protease kwaye inokwenza i-hydrolyze ezinye ii-proteases kwilebhu. Ke ngoko, i-proteinase K ifuna iindawo zemveliso ezikhethekileyo, izixhobo, kunye nabasebenzi bophando, uphuhliso kunye nemveliso.
Isisombululo se-XD BIOCHEM se-proteinase K
I-XD BIOCHEM ineqonga elivuthiweyo lokubonakalisa nokucoca iiproteni, kwaye inamava amaninzi ekubonakaliseni nasekucoceni iiproteni eziphinda zihlanganiswe kunye nokwenza ngcono iinkqubo zemveliso. Ngokusekwa ngokukhawuleza kweqela lophando nophuhliso, inkqubo enkulu yemveliso yeproteniase K iye yoyiswa. Imveliso yenyanga yomgubo owomileyo oqandisiweyo ingaphezulu kwe-30 KG. Imveliso inamandla azinzileyo, umsebenzi ophezulu we-enzyme, kwaye ayinazo ii-cytochrome kunye nee-nucleic acid residues. Wamkelekile ukuqhagamshelana ne-XD BIOCHEM Fumana iphakheji yovavanyo (I-imeyile:sales@xdbiochem.comUmnxeba: +86 513 81163739).
Izisombululo zobugcisa ze-XD BIOCHEM ziquka
Ngokusebenzisa i-multi-copy plasmid integration, kukhethwa iintlobo ze-high-expression ezine-expression level ye-8g/L, nto leyo eyoyisa ingxaki ye-low-expression level ye-proteinase K.
Ngokusekwa kwenkqubo yokucoca enezinyathelo ezininzi, i-host cytochrome kunye ne-nucleic acid residues zeproteinase K zasuswa ngempumelelo ngaphantsi kwexabiso eliqhelekileyo.
Ngokusebenzisa uhlolo oluphezulu lwe-protective buffer formulations, kukhethwe i-buffer enokugcina i-proteinase K ngokuzinzileyo kumaqondo obushushu angama-37°C.
Ii-buffers zokuhlola ziyayoyisa ingxaki yokuba kulula ukuyihlanganisa nokuyicoca i-proteinase K xa ikwizinga eliphezulu, kwaye zibeka isiseko sokomisa i-proteinase K xa ikwizinga eliphezulu le-solid content.
Isampulu ye-XD BIOCHEM proteinase K
Uvavanyo lwe-XD BIOCHEM proteinase K: akuyi kubakho tshintsho lubalulekileyo kumsebenzi emva kweeyure ezingama-80 kubushushu begumbi
Uvavanyo lwe-XD BIOCHEM proteinase K: akuyi kubakho tshintsho lubalulekileyo kumsebenzi emva kweeyure ezingama-80 kubushushu begumbi.
Ukuthelekiswa kwesiphumo sokukhupha i-nucleic acid ye-XD BIOCHEM proteinase K kunye neemveliso ezikhuphisanayo. Kwinkqubo yokukhupha i-nucleic acid, i-XD BIOCHEM kunye ne-competitive proteinase K ziyasetyenziswa ngokwahlukeneyo. Ukusebenza kakuhle kokukhupha i-XD BIOCHEM proteinase K kuphezulu kwaye ixabiso le-Ct yejini ekujoliswe kuyo liphantsi.
Ixesha lokuthumela: Disemba-31-2021