Serivisi yo gusesengura imikoranire ya poroteyine na poroteyine
Alpha Lifetech ishobora kandi guha abakiriya ibizamini bya poroteyine, isesengura ry’imikoranire ya poroteyine, isuzuma ry’imikoranire ya poroteyine na poroteyine (CO-IP, Western Blot, isuzuma rya Chromatin immunoprecipitation), n’isuzuma ry’imikorere ya poroteyine kugira ngo ihuze n’ibyo abakiriya batandukanye bakeneye mu igerageza.
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Serivisi yo gusesengura imikoranire ya poroteyine na poroteyine
Alpha Lifetech imaze imyaka myinshi ikora cyane mu isuzuma rya poroteyine, yubaka urubuga rwiza rwo gupima imikorere ya poroteyine, kandi ifite ubuhanga mu buryo butandukanye bwo kugenzura imikoranire ya poroteyine, bishobora kuzigama igihe cy'ubushakashatsi bwa siyansi cyangwa ubushakashatsi ku mishinga mu gihe bitanga umusaruro uhamye.
Alpha Lifetech ishobora kandi guha abakiriya ibizamini bya poroteyine, isesengura ry’imikoranire ya poroteyine, isuzuma ry’imikoranire ya poroteyine na poroteyine (CO-IP, Western Blot, isuzuma rya Chromatin immunoprecipitation), n’isuzuma ry’imikorere ya poroteyine kugira ngo ihuze n’ibyo abakiriya batandukanye bakeneye mu igerageza.
Intangiriro ku isuzuma ry'imikoranire ya poroteyine na poroteyine
Poroteyine ni molekile zikomeye cyane mu buzima bwose zikurikiza amabwiriza abitswe mu bikoresho bya genetike. Akenshi, poroteyine zikora akazi kazo zigakora nk'abakinnyi bagira uruhare mu gukorana n'izindi poroteyine, ibi bikaba bigaragara cyane iyo imikorere ya poroteyine isuzumwe mu buryo nyabwo bw'akaremangingo. Kumenya imikoranire hagati ya poroteyine ni ingenzi cyane mu iperereza rya biochimie kuri poroteyine imwe ku giti cyayo. Isesengura ry'imikoranire ya poroteyine mu bwoko busanzwe rikurikira:
Isuzuma rya "co-immunoprecipitation Assay"
Ihame ry'uburyo bwo gufata imiti igabanya ubukana bw'umubiri ni ukuvanga poroteyine, antibody yihariye, na poroteyine A/G magnetic beads kugira ngo icungwe, supernatant ya poroteyine idapfundikiye ijugunywa binyuze mu kwinjiza magnetic beads kuri magnetic rack, hanyuma magnetic beads zikaraba kugira ngo poroteyine n'indi myanda ikureho keretse iyo ipfundikiye ku buryo bwihariye.
Igishushanyo cya 1 cy'isuzuma rya Co-IP.(Inkomoko y'aho uherereye: Isuzuma rya Coimmunoprecipitation - PubMed (nih.gov))
Isuzuma ryo gukuraho
Isuzuma rikorwa ni ugushyira poroteyine izwi ku gasaro ka rukuruzi hanyuma wongereho ikintu kigomba kugaragara. Isuzuma riboneka kugira ngo poroteyine zifatanye zihuzwe.
Mu 1988, Smith n'abandi basukuye poroteyine ya GST fusion, yakoreshejwe mu isuzuma rya pull-down yitwa Gst pull down, ari byo kongeramo agapapuro ka GST kuri poroteyine ishakisha, gufata poroteyine ikorana binyuze muri GSH, gusobanura uko ihambiriye, no kuyigenzura binyuze mu isuzuma rya WB.
Igishushanyo cya 2 Imbonerahamwe y'ibizamini bya GST.(Inkomoko y'aho uherereye: Ikizamini cya GST cyo Kwiga uburyo PIF4 Binding In Vitro ikoreshwa - PubMed (nih.gov) )
Ikintu cy’ingenzi agamije ni ugukoresha iyoni z’icyuma nka nikeli cyangwa kobalti nk’uburyo bwo kubikoresha, gusukura poroteyine binyuze muri chromatography y’ubumwe, no gusobanura uburyo bwo gufatana binyuze mu igerageza rya WB kugira ngo harebwe niba koko ibintu bimeze.
Byongeye kandi, hari ADN Pull-down (ikizamini cyo gufatanya na poroteyine), RNA Pull-down (ikizamini cyo gukorana na poroteyine), n'ikizamini cyo gukurura molecule nto.
ADN Pull-down (isuzuma rya poroteyine dna binding assay) ni uburyo bwo kumenya ADN ifatanye na poroteyine. Niba poroteyine runaka zifatanye na ADN runaka byagaragaye hakoreshejwe isuzuma rya WB; Uduce twa ADN tutazwi dufatanye na poroteyine, uduce twa ADN tuzwi no gupimwa na MS.
Gupima RNA (Poroteyine rna interaction assay) ni uburyo bwo gusuzuma RNA ifatana na poroteyine. Niba poroteyine runaka ifatana na RNA yibasiwe byagaragaye hakoreshejwe ibizamini bya WB; Uduce twa RNA tutazwi dufatana na poroteyine, uduce twa RNA tuzwi hakoreshejwe MS.
Gupima SM (small molecule pull-down assays) bishobora kubona poroteyine zigamije gufatanya by'umwihariko na molekile nto, kandi iyo bihujwe na MS, poroteyine nto zigamije molekile zishobora gupimwa neza, zishobora kugabanywamo uburyo bwo gushyira ibirango muri vitro n'uburyo bwa orthogonal.
Igerageza rya WB
Igerageza rya WB (rizwi kandi nka Western Blot experiment cyangwa Western blot), ishingiro ni uburyo bwihariye bwa antigen na antibody, ihame ry'ibanze ni ugutandukanya poroteyine zangiritse binyuze muri electrophoresis ya SDS polyacrylamide gel no kuyishyira mu gikoresho gikomeye cyo gutwara (nk'urukiramende rwa PVDF, urukiramende rwa NC) binyuze mu kohereza urukiramende (guhinduranya amazi cyangwa se kwumye). Nyuma yo kuziba (hakoreshejwe BSA, buttermilk, nibindi), antibody y'ibanze ikoreshwa kugira ngo ifatanye by'umwihariko na poroteyine, hanyuma antibody y'inyongera ifite ikirango cya fluorescein yongerwaho nyuma yo koza agapapuro. Mu guhambira antibody ya kabiri na antibody y'ibanze, poroteyine ikoreshwa ishobora kugaragara binyuze mu iterambere ry'amabara ya substrate na chemiluminescence. Muri rusange ikoreshwa mu kumenya niba poroteyine runaka igaragara mu gipimo no gusesengura neza urwego rw'imikorere yayo.
Uburyo bwo kugereranya uburyo bwo gukorana hagati ya poroteyine na poroteyine
Isuzuma rya poroteyine rikoreshwa mu gukurura poroteyine risa n’isuzuma rya Co-IP, byombi bikaba ari ibyo gupima igerageza ry’imikoranire ya poroteyine. Itandukaniro riri hagati y’uburyo bubiri ni uko isuzuma rya Co-IP ribona poroteyine zikorana za poroteyine zizwi, zifite ukuri runaka ariko ntizishobora kwemeza niba poroteyine zikorana mu buryo butaziguye cyangwa butaziguye. Isuzuma rya poroteyine rikoreshwa mu gukurura poroteyine rigamije kubona poroteyine itazwi ikorana na poroteyine izwi. Rishobora kwemeza neza niba poroteyine izwi ikorana na poroteyine yabonetse ariko ntirishobora kumenya uko ihuza mu mubiri.
Ikoranabuhanga rya Co-immunoprecipitation rikoreshwa mu gusesengura niba hari imikoranire hagati ya poroteyine ebyiri cyangwa nyinshi, hashingiwe ku igerageza rya immunoprecipitation, Ugereranyije n'ubundi buryo bwo gusesengura imikoranire ya poroteyine mu buryo bw'igerageza (GST Pull-down, Western Blot, Chromatin immunoprecipitation assay, nibindi), ikoranabuhanga rya Co-IP rifite umwihariko wo hejuru, ubushobozi bwo kumva, no gusubiramo cyane, bishobora kwirinda kubangamirana kw'ibidahuye no kugaragaza imikoranire ya poroteyine mu buryo karemano.
| Kuramo GST hasi | Co-IP | WB | |
|---|---|---|---|
| Akamaro | *Byoroshye gukoresha *Bikwiriye gusukura poroteyine zitandukanye | *Ku isesengura ry'imikoranire mu mubiri *Ishobora kumenyekana na poroteyine iri inyuma | *Uburyo buhamye bwo kwiharira *Ubuziranenge n'ubunini mu buryo bwa "in vitro" |
| Imipaka | *Ahari guhuza ibintu bitari ngombwa *Hakenewe kwemeza byinshi ku mikoranire | *Kwishingikiriza cyane ku budahangarwa bw'umubiri *Guhuza ibintu bitari bimwe bishobora kubaho | *Bitwara igihe *Biragoye gukoresha mu gusesengura poroteyine ku rwego runini |
| Urugero rw'Ishyirwa mu Bikorwa | *Menya imikoranire *Poroteyine isukuye | *Isuzuma ry'uburyo ikimenyetso gikwirakwizwa *Imiterere y'indwara | Isesengura ryakurikiyeho ry’imikoranire ya poroteyine na poroteyine, poroteyine na ADN, n’imikoranire ya poroteyine na RNA |
Byahinduwe
Hindura serivisi kugira ngo zihuze n'ibyo ukeneye
Ubunararibonye
Uburambe bw'imyaka myinshi mu gukora poroteyine
Ikoranabuhanga
Ibisubizo by'ibitangwa bifite ireme ryo hejuru
Uburyo bwo kugerageza
Uburyo buhamye bwo gucunga ubuziranenge
Niba ufite ikibazo icyo ari cyo cyose, twandikire igihe icyo ari cyo cyose.
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2018-07-16 
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