Cov txiaj ntsig ntawm kev siv BCA khoom siv yog dab tsi?

Taw qhia rau BCA protein assay

Tus BCA (Bicinchonic acid) protein assay yog ib qho tseem ceeb ntawm cov txheej txheem ntawm biochemical thiab kev kho mob cell. Nws yooj yim ntawm cov muaj nuj nqis ntawm tag nrho cov protein ntau hauv cov qauv. Cov qauv no ua haujlwm ntawm tus qauv ntawm protein - Cov tooj liab chelation, ua raws los ntawm cov xim ua kom pom kev rau cov tooj liab uas txo tooj liab. Nws cov chaw muaj koob muaj txiaj ntsig los ntawm nws cov kev siv dav dav thoob plaws ntau cov qauv protein, suav nrog kev xaiv tshwj xeeb, ua rau cov khoom lag luam zoo rau ob qho chaw tsim khoom thiab cov chaw muag khoom hauv qhov chaw tsim khoom.

Compatibility nrog xab npum

Tuav cov qauv sib txuas

Ib qho ntawm cov ntsiab lus tseem ceeb ntawm BCA protein assay yog nws compatibility nrog xab npum. Tsis zoo li ntau lwm cov txheej txheem protein ntau, BCA assay tuaj yeem haum txog 5% los ntawm qhov piv txwv. Cov yam ntxwv no tshwj xeeb tau rau cov kws tshawb fawb thiab cov tuam ntxhab cuam tshuam nrog cov qauv nyuaj, qhov twg xab npum feem ntau siv rau cov protein ntau. Rau kev kho mob cell, qhov sib xyaw ua ke no kom muaj protein ntau txawm tias muaj cov tshuaj cuam tshuam no.

Kev ua tau zoo hauv cov xwm txheej nyuaj

Lub BCA assay muab cov txiaj ntsig kev txhim khu kev qha txawm tias muaj cov xwm txheej nyuaj. Nws lub zog tiv thaiv kev hloov pauv hauv cov protein ntau, xws li amino acid sequence sib txawv thiab sab saw, txo cov protein - rau - protein hloov pauv. Qhov kev sib txuam no yog qhov tseem ceeb rau cov neeg muab khoom tseem ceeb thiab ua kom muaj txiaj ntsig zoo rau kev tswj hwm siab rau cov protein zoo hauv cov protein.

Txo lub sijhawm teeb tsa

Efficiency nrog dilution - Cov Qauv Dawb

Cov lus qhia ntawm kev ua kom muaj kev zam - Cov qauv protein dawb hauv cov khoom siv BCA assay txo lub sijhawm teeb tsa los ntawm 80%. Cov qauv prediluted yog tsim los ntawm kev sib koom ua ke nrog cov roj multichannel pipettes, pub cov kws tshawb nrhiav thiab cov txheej txheem kom nrawm dua cov txheej txheem assay yam tsis muaj kev cuam tshuam. Rau cov ntawv kho mob cell, lub sijhawm twg - Cov kev sim rhiab yog ib qho, qhov kev ua tau zoo no muaj txiaj ntsig.

Yooj Yim Workflow

Qhov no txoj kev sib ze txo cov kev xav tau rau kev coj ua cov kauj ruam kev sib tw, uas tsis yog lub sijhawm - kev siv tab sis kuj tseem ceeb rau kev ua yuam kev. Los ntawm kev tshem tawm cov kauj ruam no, BCA assay pab nrawm nrawm, tso cai rau cov khoom xa tuaj thiab txhim kho kev tsim khoom hauv kev kuaj.

Txhim kho protein ua kom zoo nkauj

Yog qhov ntsuas tau hla cov hom protein

Lub BCA protein assay ua yeeb yam rau cov lus teb ntau ntxiv rau cov protein ntau dua rau cov chaw zas xim ntxiv xws li tus qauv Bradford. Qhov kev sib txuam no yog ua tiav los ntawm reacting proctionally rau tus naj npawb ntawm Peptide Bonds, txo cov assay txoj kev sib txawv mus rau qhov sib txawv rau cov qauv protein ntau. Rau cov tuam txhab tsim kev tsim kev tsim muaj nyob rau hauv kev tsim cell kev tshuaj xyuas, qhov no ntseeg tau qhov tseeb ntawm cov protein ntau yam protein, kev ntseeg siab hauv assay tau.

Txhim kho cov assay sib xws

Los ntawm muab cov txiaj ntsig zoo ib yam tsis hais txog cov protein ntau, tus BCA assay yog qhov tsis haum rau cov neeg xa khoom kom muaj nuj nqis rau kev tswj thiab khoom txheem. Qhov no sib xws txhais cov ntaub ntawv txhim khu kev qha ntau dua, qhov tseem ceeb rau kev tshawb fawb tshawb fawb hauv kev kho mob cell thiab lwm cov teb.

Kev rhiab thiab nrhiav tau ntau yam

High rhiab heev rau cov kev ua kom muaj ntau

Lub BCA assisy muaj qib siab ntawm kev pom zoo, muaj peev xwm ntes tau cov protein ntau li 0.5 μg, thiab tswj hwm kev ua kom txog 1.5 mg / ml. Qhov kev ntsuas dav dav no ua rau nws haum rau ntau qhov kev siv, los ntawm kev tshawb fawb yooj yim rau kev kho cov kev kho mob. Xws rhiab yog qhov tseem ceeb thaum cuam tshuam nrog feeb cov protein nyob rau hauv cell kev kho mob thiab kev muag tshuaj.

Optimal Nrhiav Kom Tau Cov Kev Ntsuas

Cov xim kuaj pom ntawm 562 nm ua kom pom tseeb tsawg teeb liab poob (tsawg dua 10%), tso cai rau cov tuam ntxhab thiab cov chaw tsim khoom kom ua tiav kom muaj nuj nqis. Qhov tseeb yog qhov tseem ceeb rau kev tsim kho cov khoom siv kho mob ntawm tes, qhov twg precise protein concentration tuaj yeem ncaj ncees cuam tshuam.

Yooj yim ntawm kev siv hauv cov txheej txheem assay

Txheej txheem streamlined

BCA assay nta tau yooj yim cov txheej txheem, muaj txiaj ntsig rau ob tus kws tshaj lij thiab cov neeg ua haujlwm lom zem. Kev npaj txhij - Siv reagents thiab ncaj ncaj raws kev ua kom muaj peev xwm ua tau, suav nrog cov neeg tsim khoom lag luam, suav nrog cov chaw tsim khoom thiab cov chaw muag khoom tshiab rau cov qauv protein.

Tus neeg siv khoom siv - tus qauv tsim

Tsim los ntawm kev siv nyob rau hauv lub siab, BCA assay minimizes kev cob qhia hurdles technology, pab txhawb kev yooj yim dua thiab nrawm dua hauv kev kuaj. Cov kev nkag tau no pab tau txoj haujlwm thoob plaws rau cov neeg muag khoom uas yuav tsum tau tuav lub siab kom tsis muaj qhov sib luag.

Qhov tseeb thiab xwm yeem

Ntseeg tau cov txiaj ntsig dhau kev sim

Lub BCA pom tias cov txiaj ntsig tau zoo, cim los ntawm cov protein tsawg - rau - protein variation. Qhov xwm txheej sib xws yog qhov tseem ceeb rau kev sim uas xav tau kev ntsuas kev ntsuas dua, xws li cov muaj protein ntau dua, qhov twg muaj cov protein ntau tshaj plaws yog qhov tseeb ntawm cov txiaj ntsig.

Cov Qauv thiab Kev Ntsuas

Cov txheej txheem tiv thaiv BSA los ntawm National Institute ntawm cov qauv thiab thev naus laus zis (nist) kom ntseeg tau tias BCA assay muab cov qauv raug kab mob. Qhov kev ntsuas hluav taws xob no tseem ceeb rau cov chaw tsim khoom thiab lwm tus neeg uas mob siab xa cov khoom siab (siab, qauv qauv.

Kev txhais tes ntawm BCA Txoj Kev

Biochemical Tshuab

Lub BCA txoj kev yog raws cov kev txo ntawm CU²⁺rau cu⁺Los ntawm cov protein nyob rau hauv ib qho nruab nrab alkaline, ua raws li los ntawm kev tsim cov ntshav ntshav nrog BCA, uas yog qhov ntsuas pom kev ua cim Coloricetric. Qhov no ob txoj kev tshaj lij ntxiv muab cov hauv paus ntseeg tau rau cov protein ntau, muaj feem xyuam hauv cov khoom tshawb fawb thiab kev lag luam kev lag luam.

Cov txiaj ntsig zoo dua lwm cov kev cia siab

Piv rau cov hau kev zoo li Bradford Assis, cov qauv BCA muaj ntau yam kev cuam tshuam nrog cov tshuaj cuam tshuam nrog cov tshuaj sib cuam tshuam nrog hom sib txawv. Qhov no ua rau nws xaiv qhov kev xaiv zoo rau cov tuam ntxhab thiab cov chaw muag khoom uas xav tau cov kev daws teeb meem zoo thiab txhim khu kev qha.

Kev sib piv nrog Bradford Assay

Qhov sib txawv tseem ceeb hauv vib this

Thaum ob qho tib si BCA thiab Bradford tseem siv tau dav siv rau cov protein kom muaj protein, lawv sib txawv hauv qab hauv lawv txoj kev. Tus Bradford cov tshuaj ntsuab kev vam khom rau cov zas - protein khi, uas tuaj yeem tsis muaj ntau yam ntawm ntau hom protein ntau. Nyob rau hauv kev sib piv, BCA assay peptide beptide boils thiab ntseeg tau muaj txiaj ntsig zoo, tshwj xeeb rau cov chaw tsim khoom thiab lwm tus neeg saib xyuas kev kho mob cell thiab cuam tshuam.

Qhov zoo ntawm BCA assay

Lub BCA assay compatibility nrog kev xab npum, cov khoom siv ua haujlwm loj dua, thiab yooj yim workamflow muab nws ib qho ntug kev xav tau cov peev xwm tshwj xeeb no. Nws cov vagatility ua rau nws haum rau tus dav dav ntawm kev sim, ib qho tseem ceeb rau cov neeg muag khoom rau cov khoom siv kev tshawb fawb.

Xaus thiab lus pom zoo

Vim li cas BCA yog qhov xav tau

Tus BCA protein assay sawv tawm rau nws cov kev sib xyaw nrog xab npum, txo lub sijhawm teeb tsa, txhim kho protein sib txawv, thiab raug. Cov cwj pwm no ua rau nws haum haum rau cov ntawv thov hauv txoj kev kho mob cell, muab cov txiaj ntsig txhim khu kev qha thiab zoo ib yam. Rau cov tuam tsev tsim khoom thiab cov chaw muag khoom, cov BCA assay muab txoj kev vam meej rau kev ua kom muaj protein ntau ntxiv, qhov tseem ceeb rau kev tswj hwm cov qauv kev lag luam thiab txhawb nqa cov khoom lag luam kev lag luam thiab cov khoom lag luam txhawb nqa.

Blukit muab cov kev daws teeb meem

Bluekit tshwj xeeb hauv kev muab cov kev kawm siab rau cov BCA protein assay, ua kom tau raws li cov kev xav tau ntawm cov kws tshawb fawb, cov chaw tsim khoom, thiab lwm tus neeg. Peb cov khoom siv tau tsim rau kev ua tau zoo, muab cov protein txhim khu kev qha thiab muaj txiaj ntsig ntau ntawm cov ntawv thov. Nrog cov bluekit, koj tuaj yeem txhim kho koj cov haujlwm ua haujlwm tau zoo thiab qhov raug, kom koj ntsib cov qauv kev kho mob ntawm cell kev tshawb fawb thiab kev tsim khoom. Cia cov khau khiab yog koj tus khub uas ntseeg siab rau tag nrho koj cov protein assay xav tau.