Qhov sib txawv tseem ceeb ntawm immunoprecipitation thiab coimmunoprecipitation yog immunoprecipitation yog ib qho txheej txheem uas precipitates cov protein tawm ntawm cov tshuaj uas siv cov tshuaj tiv thaiv tshwj xeeb, thaum coimmunoprecipitation yog cov txheej txheem uas precipitates cov protein ntau complexes tawm ntawm cov tshuaj siv cov tshuaj tiv thaiv tshwj xeeb.
Antigen-antibody kev sib cuam tshuam yog ib qho tshuaj lom neeg sib cuam tshuam ntawm cov tshuaj tiv thaiv kab mob uas tsim los ntawm B hlwb thiab antigens (proteins) thaum muaj kev tiv thaiv kab mob. Nquag, soluble antigens ua ke nrog cov tshuaj tiv thaiv soluble nyob rau hauv lub xub ntiag ntawm ib qho electrolyte ntawm soluble kub thiab pH thiaj li yuav ua rau ib tug insoluble pom complex. Qhov no yog hu ua nag lossis daus. Yog li ntawd, immunoprecipitation thiab coimmunoprecipitation yog ob hom nag lossis daus cov tshuaj tiv thaiv uas niaj hnub siv hauv chav kuaj xyuas cov proteins xws li antigens.
Immunoprecipitation yog dab tsi?
Immunoprecipitation yog ib qho txheej txheem ntawm precipitating cov protein tawm ntawm cov tshuaj siv cov tshuaj tiv thaiv tshwj xeeb. Qee zaum, cov txheej txheem no tseem hu ua tus neeg muaj protein ntau immunoprecipitation. Immunoprecipitation siv cov tshuaj tiv thaiv kab mob los cais cov protein ntau ntawm kev txaus siab los ntawm cell lysate. Cov tshuaj tiv thaiv kab mob purifies nws lub hom phiaj tshwj xeeb protein los yog antigen los ntawm kev sib tov. Cov tshuaj tiv thaiv kab mob khi rau cov protein, thiab cov tshuaj tiv thaiv-antigen complex raug rub tawm ntawm cov qauv. Hauv kev sim teeb tsa, cov antigen-antibody complex yog rub tawm siv cov protein A / G txuas nrog agarose lossis cov hlaws sib nqus. Tom qab ntawd, cov hlaws yog ntxuav, thiab cov protein ntawm kev txaus siab yog eluted. Cov purified protein los yog antigen tau los ntawm immunoprecipitation yog txheeb xyuas los ntawm ntau yam txuj ci xws li enzyme-linked immunosorbent assay (ELISA) thiab western blot (WB).
Daim duab 01: Immunoprecipitation
Tsis tas li ntawd, cais cov proteins lossis antigens tuaj yeem txheeb xyuas thiab txheeb xyuas los ntawm pawg spectrometry los ntawm kev siv cov qauv kev zom zaub mov enzymatic raws li thawj theem. Muaj qee qhov kev txiav txim siab ua ntej pib qhov kev sim tshuaj tiv thaiv kab mob: txoj kev xaiv hom, khi cov proteins thiab cov hlaws dai, xaiv tus thawj antibody thiab isotype, thiab kev tswj tsis zoo.
Coimmunoprecipitation yog dab tsi?
Coimmunoprecipitation yog ib qho txheej txheem ntawm precipitating cov protein complexes tawm ntawm cov tshuaj uas siv cov tshuaj tiv thaiv tshwj xeeb. Coimmunoprecipitation feem ntau ua haujlwm los ntawm kev xaiv cov tshuaj tiv thaiv tshwj xeeb uas tsom rau cov protein paub (antigen) uas ntseeg tau tias yog cov tswv cuab ntawm cov protein ntau dua. Los ntawm kev tsom mus rau cov tswvcuab paub nrog cov tshuaj tiv thaiv tshwj xeeb, nws muaj peev xwm rub tau tag nrho cov protein complex tawm ntawm cov tshuaj. Qhov no ua rau kev txheeb xyuas ntawm cov tswv cuab tsis paub ntawm lub complex. Lub tswv yim ntawm rub cov protein complexes tawm ntawm kev daws yog qee zaum hu ua "rub-down" mechanism.
Daim duab 02: B Coimmunoprecipitation
Coimmunoprecipitation yog cov txheej txheem muaj zog uas siv los txheeb xyuas kev sib cuam tshuam ntawm cov protein-protein. Lub hom phiaj tseem ceeb ntawm coimmunoprecipitation yog kev txheeb xyuas cov neeg koom tes sib cuam tshuam xws li ligands, cofactors, lossis cov cim qhia txog cov khoom noj rau cov protein uas txaus siab. Ntxiv mus, coimmunoprecipitation yog ib qho txheej txheem zoo uas yog siv los cais cov proteins los ntawm cov ntshav, cell lysate, homogenized cov ntaub so ntswg, lossis cov xov xwm txias.
Dab tsi zoo sib xws ntawm Immunoprecipitation thiab Coimmunoprecipitation?
- Immunoprecipitation thiab coimmunoprecipitation yog ob hom nag lossis daus-raws li cov txheej txheem.
- Ob txoj kev siv tau los txheeb xyuas lub hom phiaj antigen los ntawm cov tshuaj tiv thaiv tshwj xeeb.
- Ob txoj kev siv zog ua haujlwm nyob ntawm kev sib cuam tshuam ntawm antigen-antibody.
- Lawv siv niaj hnub siv hauv chaw kuaj mob.
Qhov txawv ntawm Immunoprecipitation thiab Coimmunoprecipitation yog dab tsi?
Immunoprecipitation yog ib qho txheej txheem uas ua kom cov protein ntau tawm ntawm cov tshuaj siv cov tshuaj tiv thaiv kab mob tshwj xeeb, thaum coimmunoprecipitation yog cov txheej txheem uas ua kom cov protein tsis zoo tawm ntawm cov tshuaj siv cov tshuaj tiv thaiv tshwj xeeb. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm immunoprecipitation thiab coimmunoprecipitation. Tsis tas li ntawd, immunoprecipitation feem ntau siv nyob rau hauv lub chaw soj nstuam, thaum coimmunoprecipitation tsis yog siv nyob rau hauv lub chaw soj nstuam.
Cov hauv qab no infographic nthuav tawm qhov sib txawv ntawm immunoprecipitation thiab coimmunoprecipitation hauv daim ntawv tabular rau ib sab ntawm kev sib piv.
Summary – Immunoprecipitation vs Coimmunoprecipitation
Immunoprecipitation thiab coimmunoprecipitation yog ob hom nag lossis daus-raws li cov tswv yim uas nyob ntawm kev sib cuam tshuam ntawm antigen-antibody. Cov txheej txheem immunoprecipitation precipitates cov protein tawm ntawm cov tshuaj uas siv cov tshuaj tiv thaiv tshwj xeeb, thaum cov txheej txheem coimmunoprecipitation precipitates cov protein complexes tawm ntawm cov tshuaj siv cov tshuaj tiv thaiv tshwj xeeb. Yog li, qhov no qhia txog qhov sib txawv ntawm immunoprecipitation thiab coimmunoprecipitation.
