Qhov sib txawv tseem ceeb ntawm colorimetric thiab fluorometric assay yog qhov kev ntsuam xyuas colorimetric txiav txim siab qhov concentration ntawm cov xim sib xyaw hauv cov tshuaj thaum fluorometric assay txiav txim siab lub kinetic mechanism ntawm kev daws.
Kev tshuaj ntsuam biochemical yog ib qho txheej txheem uas kuaj pom lossis ntsuas qhov kev ua ntawm cov tshuaj lom neeg lossis cov tshuaj analytically. Nws yog cov txheej txheem hauv vitro. Colorimetric assay thiab fluorometric assay yog ob hom kev tshuaj ntsuam biochemical uas tau ua nyob rau hauv lub chaw soj nstuam. Ntau cov tswv yim xws li ELISA thiab sab hnub poob blotting kuj yog cov tshuaj biochemical uas nyuaj rau kev txheeb xyuas cov kev ua hauv metabolic thiab ntsuas kev ua haujlwm ntawm biomolecules xws li cov proteins, enzymes, thiab lwm yam me me molecules. Cov kev ntsuam xyuas no yog siv rau kev txheeb xyuas cov protein-DNA, protein-RNA, thiab kev sib cuam tshuam ntawm cov protein-protein.
Dab tsi yog Colorimetric Assay?
Kev tshuaj ntsuam colorimetric yog cov txheej txheem uas txiav txim siab qhov concentration ntawm cov xim sib xyaw hauv cov tshuaj. Hauv lwm lo lus, colorimetric assay yog cov tshuaj tiv thaiv uas ua rau muaj kev hloov xim vim yog cov tshuaj tiv thaiv enzymatic lossis tshuaj lom neeg ntawm cov tshuaj reagents thiab cov tshuaj ntsuam xyuas. Colorimetric assay yog siv nyob rau hauv biochemistry los ntsuam xyuas cov enzymes, tej compounds, cov tshuaj hormones, antibodies thiab lwm yam analytes. Lawv siv colorimeters lossis spectrophotometers. Colorimeters yog cov cuab yeej uas qhia cov qauv xim kom muab lub hom phiaj ntsuas ntawm cov yam ntxwv xim. Lub spectrophotometer yog ib qho cuab yeej uas ntsuas lub teeb siv raws li kev ua haujlwm ntawm cov xim lossis wavelength ntawm lub teeb.
Daim duab 01: Kev Ntsuam Xyuas Xim Xim
Kev Ntsuam Xyuas Colorimetric Ua Haujlwm Li Cas?
Nyob rau hauv kev tshuaj ntsuam colorimetric, ib lub phaj yog npaj nrog ib tug tshwj xeeb antibody khi rau qhov dej. Tom qab ntawd cov qauv ntxiv. Qhov no tuaj yeem tso cai rau tus qauv khi rau cov tshuaj tiv thaiv. Tom qab ntawd cov tshuaj tiv thaiv kab mob thiab cov substrate raug ntxiv rau hauv qhov dej kom hnov mob nrog kev tshawb nrhiav. Kev daws teeb meem yog ntxiv thaum kawg ua ntej nyeem ntawv. Ib qhov khoob khoob hu ua qhov khoob yog sab laug yam tsis muaj tus qauv. Hauv kev ntsuam xyuas colorimetric, qhov xim tsaus dua, qhov ntau dua qhov kev tshuaj ntsuam xyuas. Feem ntau, tsuas yog ib lub wavelength yog xav tau los nyeem ntawv. Tab sis yog tias muaj kev ntsuas ntsuas, siv ob lossis ntau lub wavelengths.
Kev Ntsuas Fluorometric yog dab tsi?
Kev kuaj fluorometric yog ib qho txheej txheem uas txiav txim siab cov txheej txheem kinetic ntawm cov tshuaj tiv thaiv enzyme. Kev ntsuam xyuas fluorometric yuav siv qhov chaw nrog tsim cov khoom fluorescent los ntawm cov khoom tsis muaj fluorescent substrate lossis rov ua dua. Qhov kev ntsuam xyuas no tseem ua rau siv cov fluorescence resonance zog hloov pauv (FRET) qhov twg, cov tshuaj tiv thaiv enzymatic hloov txoj hauj lwm ntawm ob lub fluorophores nyob rau hauv lub substrate yog li, hloov cov fluorescence siv.
Daim duab 02: Fluorometric Assay
Fluorometric kev soj ntsuam feem ntau yog rhiab ntau dua li lwm qhov kev ntsuam xyuas. Kev kuaj mob enzyme nyob rau hauv cov ntaub so ntswg, cell, lossis cov kua dej kuaj ntawm cov neeg mob nce vim nws qhov rhiab heev.
Kev Ntsuas Fluorometric Ua Haujlwm Li Cas?
Hauv kev ntsuam xyuas fluorometric, ib qho substrate tau ntxiv rau cov qauv hauv lub phaj, thiab cov lus teb fluorescence raug coj los siv lub phaj nyeem ntawv. Ntawm no, txhua lub cell yog ntsuas nyias. Opaque daim hlau yog siv rau hauv kev tshuaj ntsuam fluorometric. Qhov no txo qhov tawg ntawm lub teeb. Ob lub wavelengths yog tsim nyog rau hom kev ntsuam xyuas no. Ib lub wavelength yog los xyuas qhov excitation thiab lwm wavelength yog rau emission.
Dab tsi zoo sib xws ntawm Colorimetric thiab Fluorometric Assay?
- Kev tshuaj ntsuam xyuas xim xim thiab tshuaj fluorometric yog ob hom kev tshuaj ntsuam biochemical.
- Ob qhov kev ntsuam xyuas tau ua tiav rau kev kuaj mob.
- Cov kev ntsuas no cuam tshuam nrog cov tshuaj tiv thaiv enzymatic.
- Ob qhov kev ntsuam xyuas suav nrog lub substrate thiab analytes.
Qhov txawv ntawm Colorimetric thiab Fluorometric Assay yog dab tsi?
Kev tshuaj ntsuam colorimetric yog ib qho txheej txheem uas txiav txim siab qhov concentration ntawm cov xim sib xyaw hauv cov tshuaj, thaum fluorometric assay yog cov txheej txheem uas txiav txim siab lub kinetic mechanism ntawm cov tshuaj tiv thaiv enzyme. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm colorimetric thiab fluorometric assay. Tsis tas li ntawd, qhov kev ntsuam xyuas fluorometric yog qhov rhiab tshaj li qhov kev ntsuam xyuas colorimetric; Yog li, fluorometric kev soj ntsuam muaj peev xwm txheeb xyuas ntau cov tshuaj ntsuam xyuas. Yog li, qhov no yog lwm qhov sib txawv tseem ceeb ntawm colorimetric thiab fluorometric assay. Tsis tas li ntawd, kev soj ntsuam fluorometric yuav tsum muaj ob lub wavelengths, thaum kev ntsuam xyuas colorimetric tau ua nrog ib lub wavelength.
Cov hauv qab no infographic tabulates ntau qhov sib txawv ntawm colorimetric thiab fluorometric assay.
Summary – Colorimetric vs Fluorometric Assay
Kev tshuaj xyuas biochemical yog txheej txheem tshuaj xyuas siv los txiav txim siab thiab ntsuas cov tshuaj tiv thaiv cellular metabolic. Colorimetric assay thiab fluorometric assay yog ob hom kev tshuaj ntsuam biochemical. Colorimetric assay yog cov tshuaj tiv thaiv uas ua rau muaj kev hloov xim vim yog cov tshuaj enzymatic lossis tshuaj lom neeg ntawm cov tshuaj reagents thiab cov tshuaj ntsuam xyuas thaum fluorometric assay yog cov txheej txheem uas siv los txiav txim lub kinetic mechanism ntawm cov tshuaj tiv thaiv enzyme. Ob qhov kev ntsuam xyuas yog nyob ntawm qhov tshuaj tiv thaiv enzymatic uas cuam tshuam nrog lub substrate thiab ib qho tshuaj ntsuam xyuas. Kev ntsuam xyuas fluorometric yog qhov rhiab ntau dua li kev tshuaj ntsuam colorimetric. Qhov tseem ceeb tshaj plaws, kev tshuaj ntsuam fluorometric yuav tsum muaj ob lub wavelengths, thaum kev tshuaj ntsuam colorimetric tuaj yeem ua tiav nrog tsuas yog ib lub wavelength. Yog li, qhov no qhia txog qhov sib txawv ntawm colorimetric thiab fluorometric assay.
