Yiziphi izinzuzo zokusebenzisa i-BCA Kit?

Isingeniso se-BCA Protein Assay

I-BCA (Bicinchoninic Acid) Protein Assay iyindlela ebalulekile yokuhlaziya endaweni yocwaningo lwe-biochemical kanye nokwelashwa kweseli. Kusiza ukuncishiswa kwenani eliphelele leprotheni ngesampula. Le ndlela isebenza ngomgomo wamaprotheni - I-Chelation Copper, elandelwa ukutholwa kombala okunciphisiwe kwe-ions yethusi okuncishisiwe. Ukuthandwa kwayo kuvela ekusebenzeni okubanzi kwalo kuwo wonke amasampula amaprotheni amaningi, kufaka phakathi izengezo ezifana nezingqinamba, okwenza kube yisinqumo esithandwayo sabo bobabili abakhiqizi nabathengisi ensimini.

Ukuhambisana nama-detergents

Ukuphatha amasampula ayinkimbinkimbi

Enye yezinzuzo eziphawuleka ze-BCA Protein assuray ukuhambisana kwako okuhambisana nama-detergents. Ngokungafani nezinye izindlela eziningi zamaprotheni, i-BCA Assay ingahlalisa kuze kufike ku-5% ama-surfactants ngaphakathi kwesampula. Lesi sici sizuzisa ikakhulukazi abacwaningi futhi abakhiqizi babhekana namasampula ayinkimbinkimbi, lapho ama-detergents avame ukusetshenziselwa ukusola amaprotheni. Okwezinhlelo zokusebenza zokwelashwa kwamaselula, lokhu kuvumelana kuqinisekisa ukuthi ukunqunyelwa kwamaphrotheni kuhlala kunembile yize kukhona lezi zinto eziphazamisayo.

Ukusebenza Ezimweni Eziyinselele

I-BCA Assay ihlinzeka ngemiphumela ethembekile ngisho nasezimweni eziyinselele zokuhlola. Ukuqina kwayo ngokumelene nokwehluka kokuhlanganiswa kwamaphrotheni, njengokungafani kwe-amino acid kanye namaketanga aseceleni, kunciphisa amaprotheni - ukuhluka kweprotini. Lokhu kufana kubalulekile kubahlinzeki abadinga imiphumela engaguquki futhi evuselelwa kabusha ukuze kugcinwe okuphezulu - Izindinganiso zekhwalithi ezibonelweni zamaprotheni.

Isikhathi sokusetha setha

Ukusebenza kahle nge-dilution - amazinga wamahhala

Ukwethulwa kwe-dillotion - Amazinga wamaprotheni wamahhala e-BCA Assay Kit kunciphisa kakhulu isikhathi sokusetha esenziwa ngama-80%. Lezi zindinganiso ezivelele zenzelwe ukuhlanganiswa okungenamthungo nama-Pipette we-multichannel, avumele abacwaningi kanye nabakhiqizi ukuthi basheshise inqubo ye-Assay ngaphandle kokubekezelela ukunemba. Okwezicelo zokwelashwa kwamaselula, lapho isikhathi - izivivinyo ezibucayi zivamile, lokhu kusebenza kubalulekile.

Ukuhamba komsebenzi okwenziwe lula

Le ndlela ehleliwe inciphisa isidingo sezinyathelo ze-dilution ebhukwayo, okungeyona ukuphela kwesikhathi - ukudla kepha futhi kuthambekele emaphutheni. Ngokuqeda lezi zinyathelo, i-BCA Assay isiza ukuguquguquka okusheshayo, okuvumela abahlinzeki ukuthi bahlangabezane namashejuli wokulethwa kwezidingo futhi bathuthukise umkhiqizo ngezilungiselelo zelebhu.

Ukufana kwamaprotheni athuthukisiwe

Ukulinganisa okunembile ezinhlotsheni zamaprotheni

I-BCA Protein Assay ikhombisa impendulo efana nomfaniswano kuma-protein ahlukene uma kuqhathaniswa nodayi - Izinjongo Zokubopha njenge-Bradford. Lokhu kufana kutholakala ngokuphendula ngokulingana nenombolo yama-porttide amabhondi, ukunciphisa ukuzwela kwe-Assay kumahluko ezinhlakeni zamaprotheni. Kwabakhiqizi ababambe iqhaza ekukhiqizeni ukwelashwa kwamaselula, lokhu kuqinisekisa ukukalwa okunembile kulo lonke ulwazi lweprotheni ehlukahlukene, kuqinisa ukuzethemba ngemiphumela yokucela.

Ngcono ukuvumelanisa

Ngokuhlinzeka ngemiphumela engaguquki kungakhathalekile ukwehluka kwamaprotheni, i-BCA Assay ibaluleke kakhulu kubahlinzeki abadinga ukucaciswa okuqondile kokulawulwa kwekhwalithi kanye nokulinganiswa komkhiqizo. Lokhu kungaguquguquki kuhumushela kwidatha ethembekile ngokwengeziwe, kubalulekile ukuze kuthuthukiswe ucwaningo ekwelashweni kwamaseli kanye nezinkundla ezihlobene.

Uhla lokuzwela nokwatholwa

Ukuzwela okuphezulu kokugxila okuphansi

I-BCA ASSAY inikeza izinga eliphakeme lokuzwela, elikwazi ukuthola ukugxila kwamaprotheni aphansi njenge-0.5 μg / ml, futhi ligcina ulayini kuze kufike ku-1.5 mg / ml. Le range ebanzi ebanzi yenza ilungele izinhlelo ezahlukahlukene, kusuka ocwaningweni oluyisisekelo kuya ekuthuthukisweni kwezokwelapha okuthuthukile. Ukuzwela okunjalo kubalulekile lapho ubhekene nenani lemizuzu yamaprotheni ekwelashweni kweseli kanye nocwaningo lwekhemisi.

Amapharamitha atholakalayo afanele

Ukutholwa kwe-colorimetric ku-562 NM kuqinisekisa ukulahleka kwesiginali okuncane (okungaphansi kwe-10%), okuvumela abakhiqizi kanye nabathengisi ukufezekisa ukulinganiswa okuqondile. Lokhu kunemba kubalulekile ekuthuthukiseni imikhiqizo yokwelashwa kwamaselula ephumelelayo, lapho ukugxishwa okuqondile kwamaprotheni kungathinta ngokuqondile ukusebenza kahle kwezokwelapha.

Ukusebenziseka kalula ezinhlosweni ze-Assay

Izinqubo ezihleliwe

I-BCA Assay ifaka izinqubo ezenziwe lula, inenzuzo kubo bobabili abasebenzisi abasha kanye nabachwepheshe abanolwazi. Ukususelwa ku- - ukusebenzisa ama-reagents kanye ne-protocol eqondile kunciphisa amaphutha okungenzeka, okwenza kwafinyeleleka ezilalelini ezibanzi, kufaka phakathi abakhiqizi kanye nabathengisi abasha ngezindlela ze-protein.

Umsebenzisi - umklamo onobungane

Yakhelwe kalula ukusetshenziswa engqondweni, i-BCA Assay inciphisa izithiyo zobuchwepheshe, yenza lula ukuqeqeshwa okulula nokwamukelwa ngokushesha ngaphakathi kwezilungiselelo zelebhu. Lokhu kufinyeleleka kusiza ukusebenza kokuqondisa kwabahlinzeki abadinga ukugcina ukugcwala okuphezulu ngaphandle kokuyekethisa ukunemba.

Ukunemba nokungaguquguquki

Imiphumela ethembekile kuzo zonke izivivinyo

I-BCA ASSAy ihambisa imiphumela ethembekile, ibonakaliswa ngamaprotheni aphansi - Ku- - Ukuhluka kwe-Protein. Lokhu kuvumelana kubalulekile ukuthi izivivinyo zidinga ukulinganiswa okuphindaphindiwe ngokuhamba kwesikhathi, njengalezo ezisezelweni zokwelashwa kwamaselula, lapho ukunqunyelwa okunembile kwamaprotheni kufana nokuzala kanye nokuqinisekiswa kwemiphumela.

Ukuma nokulinganisa

Ukulinganisa okuphikisana ne-BSA ehlanzekile kusuka kuNational Institute of Standards and Technology (NITS) kuqinisekisa ukuthi i-BCA Assay inikeza ama-curve ajwayelekile futhi azenzekayo. Lokhu kulinganiswa kubalulekile kubakhiqizi kanye nabahlinzeki abahlinzeki kuhlose ukuletha okuphezulu - Ikhwalithi, Imikhiqizo Emilelwe.

Izici zobuchwepheshe zendlela ye-BCA

Inqubo ye-biochemical

Indlela ye-BCA isuselwa ekunciphiseni kwe-CU²⁺ku-cu⁺Ngamaprotheni aphakathi nendawo, elandelwa ukwakheka kwenkimbinkimbi onsomi nge-BCA, esebenza njengenkomba yombala. Lokhu okubili - Ukusabela kwesinyathelo kunikeza isisekelo esithembekile sokuqanjwa kwamaprotheni, okusebenzayo ocwaningweni oluhlukahlukene kanye nezimo zezimboni, kufaka phakathi ukuthuthukiswa komkhiqizo weselula.

Izinzuzo ngaphezu kwezinye izindawo

Uma kuqhathaniswa nezindlela ezinjenge-Bradford Assay, indlela ye-BCA inikezela ngezindlela eziningi eziguquguqukayo ngenxa yokuhambisana kwayo okubanzi ngezinto zokuphazamisa kanye nokusebenza kwayo okuguqukayo kwayo ezinhlotsheni ezihlukile zamaprotheni. Lokhu kwenza kube ukukhetha okuthandwayo kwabakhiqizi nabahlinzeki abadinga izixazululo zokuguquguquka okuguquguqukayo nezethembekile.

Qhathanisa neBradford Assay

Umehluko osemqoka ngendlela yezendlela

Ngenkathi zombili izindlela ze-BCA neBradford zisetshenziselwa kabanzi ngama-protein aveteation, ziyehluka ngokuyisisekelo ezindaweni zazo. I-Bradford Assay ithembela udayi: Ukubopha kwamaprotheni, okungaguquguquki kancane ezinhlotsheni ezahlukahlukene zamaprotheni. Ngokuphambene nalokho, isibopho se-BCA Assay's Peptide - Ukutholwa okusekelwe kunikeza ukufana okukhulu kanye nokwethenjwa, okuwusizo ikakhulukazi kubakhiqizi nabahlinzeki abagxile ekusweleni kwamaselula kanye nemikhakha ehlobene.

Izinzuzo ze-BCA Assay

Ukuhambisana kwe-BCA assay's nge-detergents, ibanga elibanzi elinamandla, kanye nokuhamba komsebenzi okwenziwe lula kunikeze umphetho wezinhlelo zokusebenza ezidinga la makhono athile. Ukusebenza kwayo kwaso kwenza kufanelekile ukuthola uhlu olubanzi lwezilungiselelo zokuhlola, isici esibucayi salabo abahlinzeki abahlinzeka ngemikhiqizo yezidingo ezahlukahlukene zocwaningo.

Isiphetho Nezincomo

Kungani kukhethwa i-BCA

I-BCA Protein Assay imele ukuhambisana kwayo ngama-detergents, inciphise isikhathi sokusetha, ukuthuthukiswa okuthuthukile kwamaprotheni, nokunemba. Lezi zimfanelo zenza ukuthi ilungele kakhulu izinhlelo zokusebenza ekwelashweni kwamaseli, inikeza imiphumela ethembekile futhi engaguquki. Kwabakhiqizi nabaphakeli, i-BCA Assay ihlinzeka ngendlela ethembekile yokuqinisekisa ukuphakama - Ikhwalithi ye-Protein yekhwalithi, ebalulekile yokugcina amazinga embonini kanye nokusekela ukuthuthukiswa komkhiqizo.

I-Bluekit ihlinzeka ngezixazululo

I-Bluekit isebenza ngokunikeza izixazululo ezithuthukile zeprotheni ye-BCA assuray, ehambisana nokuhlangabezana nezidingo ezithile zabaphenyi, abakhiqizi, kanye nabahlinzeki. Ama-kits ethu enzelwe ukusebenza okwenziwe kahle, ukuhlinzeka ngenqubekela phambili ethembekile nengaguquguquki yamaprotheni ezinhlobonhlobo zezicelo. Nge-BlueKit, ungathuthukisa ukusebenza kahle kokuhamba komsebenzi kanye nokunemba, ukuqinisekisa ukuthi uhlangabezana namazinga aphezulu kakhulu eCelle Therapy Research kanye nentuthuko yomkhiqizo. Vumela i-Bluekit ibe ngumlingani wakho othembekile kuzo zonke izidingo zakho ze-protein assay.