Kodi maubwino ogwiritsa ntchito BCA ndi ati?

Mafala Akutoma BCA Protein Antay

BCA (bicinchoninic acid) Destein Assin ndi njira yofunika yowunikira mu remochemical kafukufuku wa biocy. Imathandizira kuchuluka kwa mapuloteni onse mu zitsanzo. Njirayi imagwirira ntchito pamalingaliro a mapuloteni - kunyengerera zamkuwa, kutsatiridwa ndi mtundu wa utoto wochepetsedwa. Kutchuka kwake kumayambira kuchokera pakugwira ntchito kwake kwakukulu mu zitsanzo zamapuloteni, kuphatikizapo omwe ali ndi zowonjezera monga zowonjezera, ndikupangitsa kuti akhale chisankho kwa opanga onse ndi othandizira m'munda.

Kugwirizana ndi zotchinga

Kuthana ndi zitsanzo zovuta

Chimodzi mwazabwino za BCA mapuloteni ndi kulumikizana kwake ndi zotchinga. Mosiyana ndi njira zina zambiri zoyeserera, bcass imatha kukhala mpaka 5% zowonjezera mkati mwa zitsanzo. Izi ndizopindulitsa makamaka kwa ofufuza komanso opanga zomwe akuchita ndi zitsanzo zovuta, pomwe zopatsira mipando nthawi zambiri zimagwiritsidwa ntchito posungira mapuloteteni. Mwa kugwiritsa ntchito kwa maselo, kulumikizana kumeneku kumatsimikizira kuti mapuloteni opanga mapuloteni amakhalabe olondola ngakhale kuti pali zinthu zomwe zimasokoneza zinthu izi.

Magwiridwe antchito ovuta

A BCA atcheray imapereka zotsatira zabwino ngakhale m'mavuto oyesera. Kulimba kwake motsutsana ndi kapangidwe ka mapuloteni, ngati amino acid mitetions kusiyana ndi maunyolo ammbali, amachepetsa mapuloteni - Ma protein Kufananaku ndikofunikira kwa othandizira omwe amafunikira zotsatira zosasinthika komanso zowonjezera kukhalabe - Miyezo yapamwamba mu mapuloteni anthawi.

Kuchepetsa nthawi

Kuchita bwino ndi kuchepetsedwa - Miyezo yaulere

Kukhazikitsidwa kwa kuchepetsedwa - Miyezo ya protein yaulere mu BCA Assay Kit amachepetsa nthawi yokhazikitsa mpaka 80%. Miyezo yodziwikika iyi idapangidwa kuti ikhale yophatikizika yophatikizika ndi ma piacetes a multicnnel, ololeza ofufuza ndi opanga kuti athetse njirayo popanda kunyalanyaza kulondola. Kwa magwiridwe antchito am'madzi, nthawi - zoyeserera zofananira ndizofala, izi ndizothandiza.

Ntchito Yosavuta

Njira yolowera ikuchepetsa kufunikira kwa njira zamagalimoto, zomwe sizokhazo zokha - zotakamwa komanso zimakonda zolakwa. Pochotsa masitepe awa, a BCA akuthandizira potembenuka mwachangu, kulola othandizira kuti akwaniritse madongosolo owonjezera ndikuwonjezera zokolola m'makonzedwe a labotale.

Onjezerani ma protein

Kukhazikika kolondola pamitundu yama protein

Ascatein Assein amawonetsa kuyankha kwa yunifolomu yambiri yoyerekeza ndi matoma - kumanga njira ngati njira yokhazikika. Kufanana uku kumatheka poyankha ndalama za zowunikira za Peptide, kuchepetsa chidwi cha malingaliro a malingaliro a malingaliro a posankha m'magulu mapuloteni. Kwa opanga zopanga ma cell, izi zimatsimikizira kuti muyezo woyenera m'mitundu yosiyanasiyana ya mapuloteni, imakulitsa chidaliro pakugamula.

Kusintha kwamphamvu

Mwa kupereka zotsatira zosasinthika mosasamala mapuloteni osiyanasiyana, bcassion ndi othandiza kwa ogulitsa omwe amafunikira kuchuluka koyenera kuwongolera ndi kugwirizanitsa matenda. Kusasinthika kumeneku kumamasulira kudalirika kodalirika, ndikofunikira pakukonzekera kafukufuku wama cell komanso minda yofananira.

Chidwi ndi kudziwika

Chidwi chachikulu cha otsika otsika

The BCA Assay imapereka chidwi chachikulu, chomwe chimatha kudziwa mapuloteni otsika ngati 0,5 μg / ml, ndikusunga mzere mpaka 1.5 ml. Mitundu yotakatayi imapangitsa kuti ikhale yoyenera pamapulogalamu osiyanasiyana, kuyambira kafukufuku woyambira kupititsa patsogolo achire. Kukhuta kotereku ndikofunikira pochita ndi mapuloteni ochepa mu cell mankhwala ochizira ndi kafukufuku wamankhwala.

Magawo owoneka bwino

Kuzindikira kwa utotometric pa 562 nm kumapangitsa kutayika kocheperako (kochepera 10%), kulola opanga ndi othandizira kuti akwaniritse zochulukirapo. Kulondola kumeneku ndikofunikira pakupanga zinthu zothandiza foni ndi ma cell, komwe kukhazikika kwa kampani kumatha kusintha mwachindunji.

Kugwiritsa Ntchito Kugwiritsa Ntchito Mwa Njira Yakaunti

Njira zochepetsera

Buku la BCA likufanana ndi njira zosavuta, zopindulitsa kwa ogwiritsa ntchito atsopano ndi akatswiri opanga. Wokonzeka - - - Protocol yolumikizidwa ndi Protocol imachepetsa zolakwika, zomwe zimapangitsa kuti zikhale zolakwa zazikulu, kuphatikiza ndi opanga ndi othandizira atsopano a protein kuchuluka kwa protein.

Wosuta - Kapangidwe kabwino

Zopangidwa momasuka mosavuta, BCA ikuchepetsa zovuta zaukadaulo, kuwongolera mosavuta maphunziro komanso kukhazikitsidwa mwachangu mkati mwa makonda a labotale. Izi zimathandizanso kugwiritsa ntchito magwiridwe antchito othandizira omwe amafunikira kukhala ndi zipatso zambiri popanda kunyalanyaza kulondola.

Kulondola komanso kusasinthika

Zotsatira Zodalirika Pansi pa Zoyeserera

BCA ikugwira ntchito zodalirika, zodziwika ndi mapuloteni otsika - to - kusiyanasiyana kwa mapuloteni. Kusasinthika kumeneku ndikofunikira pakuyesera kofunikira muyeso nthawi, monga omwe amathandizira ma cell, kumene kuchuluka kwa kampani yolondola ndi kofunikira pakubala komanso kutsimikizika kwa zotsatira zake.

Kukhazikika ndi kutchuka

Kukhazikika kwa Oyeretsa BSA kuchokera ku National Institute pamlingo ndi ukadaulo (Nist) amatsimikizira kuti BCA Assion imapereka ma curve olondola komanso opangidwa bwino. Kazembeyu ndi wofunikira kwa opanga ndi othandizira omwe akufuna kupereka - zabwino, zoyenerera.

Zojambula zaluso za BCA

Njira yazomwe imachitika

Njira ya BCA imakhazikitsidwa pa kuchepetsa kwa cu²⁺kwa cu⁺ndi mapuloteni mu sing'anga yapafupi, yotsatiridwa ndi mapangidwe ofiira ndi BCA, omwe amasunga chizindikiro cha utotometric. Awiri awa - Zochita izi zimapereka maziko odalirika opanga mapuloteni, omwe amagwiritsidwa ntchito mu kafukufuku wosiyanasiyana ndi mikhalidwe yopanga mafakitale, kuphatikizapo ma cell mankhwala.

Ubwino Woposa Maganizo Ena

Poyerekeza ndi njira ngati bradford, njira ya BCA imaperekanso patsogolo kwambiri chifukwa chogwirizana ndi zinthu zomwe zimapangitsa kuti pakhale mitundu yosiyanasiyana ya mapuloteni. Izi zimapangitsa kuti ikhale chisankho chopanga opanga ndi othandizira omwe amafunikira zosintha kapena zodalirika zofuna za protein.

Kuyerekeza ndi Bradford Atay

Kusiyana kwakukulu mu njira

Pomwe onse a BLA ndi Bradford amagwiritsidwa ntchito kwambiri chifukwa cha mapuloteni opanga mapuloteni, amasiyana makamaka munjira zawo. A Bradford Assies amadalira - mapuloteni akumanga, omwe amakhala osasinthika m'mitundu yosiyanasiyana yama protein. Mosiyana ndi izi, kuphatikizika kwa bcay - Kuzindikira kwa BCA

Zabwino za bcay

Kugwirizana kwa BCA ndikuphatikizika, yowonjezera mphamvu yamphamvu, komanso ntchito yosavuta ipatseni m'mphepete pazogwiritsa ntchito zomwe zikufunika. Kusintha kwake kumapangitsa kuti zikhale zoyenera zoyeserera zoyesera, chinthu chovuta kwambiri kwa ogulitsa omwe amapereka zofunikira zofufuzira zosiyanasiyana.

Pomaliza ndi Malangizo

Chifukwa chiyani BCA imakonda

Kuyesa kwa BCA protein kumayimitsidwa kuti azigwirizana ndi zotupa zake, nthawi yochepetsetsa, yowonjezera ma protein ofananira, komanso olondola. Makhalidwe awa amapangitsa kukhala koyenera kwambiri pakugwiritsa ntchito kwa cell, kupereka zotsatira zodalirika komanso zotsatira mosasinthasintha. Kwa opanga ndi othandizira, Bullints a BCA imapereka njira yodalirika yotsimikizira - zoyeserera zabwino, ndizofunikira pakukhalabe miyezo ya mafakitale komanso kuthandiza chitukuko chazogulitsa.

BlueKKit Njira

Bluekit imathandizira kupeza mayankho apamwamba a BCA Protein Caltory, yolumikizidwa kuti ikwaniritse zosowa zenizeni za ofufuza, opanga, ndi othandizira. Mafumu athu amapangidwira kuti agwiritsidwe ntchito, kupereka zolimbitsa thupi zodalirika komanso zosakhazikika pamapulogalamu osiyanasiyana. Ndi Bluekit, mutha kukulitsa ntchito yanu yofungizira komanso kulondola kwanu, ndikuonetsetsa kuti mukumana ndi miyezo yapamwamba kwambiri mu kafukufuku wa cell ndi kukula kwa mankhwala. Lolani wabuluko kukhala mnzanu wodalirika pazosowa zanu zonse za mapuloteni.