Qhov sib txawv tseem ceeb ntawm bradford thiab lowry protein assay yog Bradford protein assay yog raws li qhov absorbance hloov ntawm cov xim Coomassie ci ntsa iab G-250 thaum Lowry protein assay yog raws li cov tshuaj tiv thaiv ntawm tooj liab ions (Cu +) ions. Ua los ntawm oxidation ntawm peptide bonds nrog Folin-Cioc alteu reagent.
Kev ntsuam xyuas yog cov txheej txheem tshuaj ntsuam xyuas uas pab qhia cov yam ntxwv tseem ceeb ntawm cov qauv. Yog li ntawd, qhov kev ntsuam xyuas tuaj yeem yog qhov kev xeem zoo lossis ntau yam. Ntau qhov chaw suav nrog kev kuaj tshuaj, kws tshuaj, ib puag ncig biology, molecular biology, biochemistry, thiab kev tiv thaiv kab mob siv cov kev ntsuas no niaj hnub. Bradford thiab Lowry protein assay yog ob qho kev ntsuam xyuas biochemical uas txiav txim siab cov protein ntau hauv cov qauv kev daws teeb meem. Ob qhov kev ntsuam xyuas siv cov txheej txheem colorimetric los muab cov txiaj ntsig.
Bradford Protein Assay yog dab tsi?
Bradford protein assay yog cov txheej txheem spectroscopic analytical ceev rau kev tsom xam cov protein. Nws qhia tau hais tias muaj tseeb siab thaum ntsuas cov protein concentration hauv cov tshuaj. Marion Bradford tau qhia txog cov txheej txheem no hauv xyoo 1976. Hauv qhov kev ntsuam xyuas no, tag nrho cov tshuaj tiv thaiv yog raws li cov amino acid muaj pes tsawg leeg ntawm cov proteins ntsuas. Nyob rau hauv lwm yam lus, Bradford protein assay yog ib tug colorimetric assay. Nws siv cov dye Coomassie ci ntsa iab xiav. Yog li ntawd, qhov kev ntsuam xyuas colorimetric protein nyob ntawm qhov kev hloov pauv ntawm cov xim. Coomassie ci ntsa iab G-250 muaj nyob rau hauv peb hom: cationic (liab), anionic (xiav) thiab nruab nrab (ntsuab). Thaum lub sij hawm acidic, daim ntawv liab ntawm cov xim tau hloov mus rau xiav. Nws lees paub qhov kev sib txuas ntawm cov protein. Yog tias tsis muaj protein ntau, cov tshuaj yuav nyob twj ywm hauv xim av.
Daim duab 01: Bradford protein assay
Bradford protein kev soj ntsuam txawv ntawm lwm qhov kev ntsuam xyuas cov protein vim nws tsis tshua muaj kev cuam tshuam ntawm ntau yam tshuaj lom neeg muaj nyob hauv cov kua protein. Cov tebchaw no suav nrog sodium, potassium, qabzib, thiab sucrose, thiab lwm yam.
Lowry Protein Assay yog dab tsi?
Lowry protein assay yog ib qho kev ntsuam xyuas biochemical siv los txiav txim tag nrho cov qib protein hauv cov tshuaj. Oliver H. Lowry yog tus neeg uas tsim cov reagent no hauv xyoo 1940. Cov txheej txheem qhia txog tag nrho cov protein concentration ntawm cov tshuaj los ntawm kev hloov xim uas yog proportional rau cov protein concentration hauv cov tshuaj. Li no, qhov no kuj yog ib qho kev ntsuam xyuas colorimetric protein.
Cov tshuaj tiv thaiv ntawm tooj liab ions (Cu +) tsim los ntawm oxidation ntawm peptide bonds thiab Folin-Cioc alteu reagent yog lub hauv paus ntawm txoj haujlwm no. Tsis tas li ntawd, cov reagent no muaj phosphomolybdic acid thiab phosphor-tungstic acid. Txawm li cas los xij, cov tshuaj tiv thaiv ntawm Lowry protein assay tsis to taub zoo. Tab sis nws koom nrog oxidation ntawm cysteine, tryptophan thiab tyrosine residues los ntawm kev txo cov Folin-Cioc alteu reagent.
Daim duab 02: Lowry protein assay
Cov cysteine residues pab txhawb rau lub absorbance pom nyob rau hauv Lowry protein assay thiab cov tshuaj tiv thaiv ua rau ib tug ci ntsa iab molecule xiav; heteropoly molybdenum xiav. Qhov txo qis ntawm cov molecule no yog ntsuas los ntawm absorbance ntawm 660nm. Li no, qhov concentration ntawm cysteine thiab tryptophan residues uas txo Folin-Cioc alteu reagent deduces tag nrho cov concentration ntawm cov protein nyob rau hauv cov tshuaj.
Dab tsi zoo sib xws ntawm Bradford thiab Lowry Protein Assay?
- Bradford thiab Lowry cov tshuaj ntsuam xyuas protein txiav txim siab cov protein ntau hauv cov tshuaj.
- Ob txoj kev yog txoj kev colourimetric.
- Kuj, qhov kev nkag siab ntawm ob txoj hauv kev yog siab.
Qhov txawv ntawm Bradford thiab Lowry Protein Assay yog dab tsi?
Bradford thiab Lowry protein assay yog ob hom kev ntsuam xyuas uas txiav txim siab cov protein ntau hauv cov tshuaj. Bradford protein assay cia siab rau qhov absorbance hloov ntawm cov xim Coomassie ci ntsa iab xiav G-250 thaum Lowry protein kev soj ntsuam nyob ntawm cov tshuaj tiv thaiv ntawm tooj liab ions tsim los ntawm oxidation ntawm peptide bonds, nrog Folin-Cioc alteu reagent. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm Bradford thiab Lowry protein kuaj. Bradford protein kev soj ntsuam yuav siv sij hawm 15 feeb los tsim qhov tshwm sim thaum Lowery protein kev soj ntsuam yuav siv sij hawm 40-60 feeb los tsim qhov tshwm sim. Yog li, qhov no yog lwm qhov sib txawv ntawm Bradford thiab Lowry protein assay. Tsis tas li ntawd, Bradford protein assay yog nyob ntawm cov amino acid muaj pes tsawg leeg thaum Lowry protein assay yog ib feem nyob ntawm cov amino acid muaj pes tsawg leeg. Li no, qhov no kuj yog qhov txawv ntawm Bradford thiab Lowry protein assay.
Hauv qab no inforaphic muab cov ntsiab lus ntxiv ntawm qhov sib txawv ntawm Bradford thiab Lowry protein assay.
Summary – Bradford vs Lowry Protein Assay
Kev ntsuam xyuas yog ib qho kev tshuaj ntsuam xyuas siv los qhia txog qhov tseem ceeb ntawm cov qauv. Bradford thiab Lowry protein assay yog ob hom kev ntsuam xyuas cov protein uas ua haujlwm raws li cov txheej txheem colorimetric. Bradford thiab Lowry protein kev soj ntsuam xyuas cov protein ntau hauv cov tshuaj. Txawm li cas los xij, qhov sib txawv tseem ceeb ntawm Bradford thiab Lowry Protein Assay nyob ntawm cov txheej txheem colorimetric lawv siv. Bradford protein assay siv Coomassie ci ntsa iab xiav G-250 thaum Lowry protein assay siv tooj liab ions (Cu +) ions thiab Folin-Cioc alteu reagent. Tsis tas li ntawd, txoj kev Bradford muab cov txiaj ntsig sai dua li Lowry protein kuaj. Txawm li cas los xij, ob txoj hauv kev no yog cov txheej txheem rhiab heev thiab raug cuam tshuam los ntawm ntau yam khoom.
