Polyacrylamide Gel Electrophoresis
Gel electrophoresis inzira yakakosha mumarabhoritari mukati meiyo biological dzidziso, inobvumira kupatsanurwa kwemacromolecules akadai seDNA, RNA nemapuroteni. Midhiya yekuparadzanisa yakasiyana uye nzira dzinobvumira zvikamu zvemamorekuru aya kuti zviparadzaniswe zvakanyanya nekushandisa maitiro avo emuviri. Kune mapuroteni kunyanya, polyacrylamide gel electrophoresis (PEJI) kazhinji ndiyo nzira yesarudzo.
PEJI inzira inopatsanura macromolecules akadai semapuroteni zvichienderana nekufamba kwawo kwe electrophoretic, ndiko kuti, kugona kwevanoongorora kuenda kune electrode yeinopesana nechaji. MuPEJI, izvi zvinotemerwa nechaji, saizi (morekuru huremu) uye chimiro chemorekuru. Vanoongorora vanofamba nemumapores akaumbwa mupolyacrylamide gel. Kusiyana neDNA neRNA, mapuroteni anosiyana mukubhadharisa zvinoenderana neamino acids akabatanidzwa, ayo anogona kukanganisa mafambiro aanoita. Tambo dzeamino acid dzinogonawo kugadzira zvimiro zvechipiri zvinokanganisa hukuru hwadzo uye nekudaro mafambisirwo avanoita nemapores. Naizvozvo dzimwe nguva zvingave zvinodiwa kuti denature mapuroteni pamberi pe electrophoresis kuti mutsara iwo kana fungidziro yakanyatsojeka yehukuru ichidikanwa.
SDS PEJI
Sodium-dodecyl sulfate polyacrylamide gel electrophoresis inyanzvi inoshandiswa kupatsanura mapuroteni mamorekuru ehuwandu 5 kusvika 250 kDa. Mapuroteni akapatsanurwa chete pahwaro hwehuremu hwema molecular. Sodium dodecyl sulfate, anionic surfactant, inowedzerwa mukugadzirira kwemajeri anovharidzira machaji emukati emapuroteni samples uye achivapa mutero wakafanana kune huwandu hwehuwandu. Nemashoko akareruka, inodenha mapuroteni uye inovapa mhosva yakaipa.
Native PEJI
Native PEJI inyanzvi inoshandisa gel dzisina-denatured yekuparadzanisa mapuroteni. Kusiyana neSDS PEJI, hapana denaturing agent inowedzerwa mukugadzirira kwemajeri. Somugumisiro, kupatsanurwa kwemapuroteni kunoitika pahwaro hwekubhadhara uye ukuru hwemapuroteni. Muhunyanzvi uhu, kuwirirana, kupeta uye amino acid cheni dzemapuroteni ndizvo zvinhu izvo kupatsanurwa kunoenderana nazvo. Mapuroteni haana kukuvadzwa mukuita uku, uye anogona kudzoserwa mushure mekupedzwa kwekuparadzana.
Ko polyacrylamide gel electrophoresis (PEJI) inoshanda sei?
Nheyo yakakosha yePEJI ndeyekuparadzanisa vaongorori nekuvapfuura nemumapores epolyacrylamide gel uchishandisa magetsi. Kuti izvi zviitike, musanganiswa we acrylamide-bisacrylamide unoitwa polymerized (polyacrylamide) nekuwedzera ammonium persulfate (APS). Mhinduro, iyo inogadziriswa ne tetramethylethylenediamine (TEMED), inoumba mambure akafanana nemaitiro ane pores ayo anoongorora anogona kufamba (Mufananidzo 2). Iyo yakakwira chikamu cheiyo acrylamide yakazara inosanganisirwa mujeri, iyo idiki saizi yepore, saka madiki mapuroteni anozokwanisa kupfuura. Huyero hwe acrylamide kune bisacrylamide huchabatawo saizi yepore asi izvi zvinowanzo chengetwa. Madiki madiki pore saizi zvakare anoderedza kumhanya uko mapuroteni madiki anokwanisa kufamba nemujeri, achivandudza kugadzirisa kwawo uye kudzivirira kuti asamhanyire mubuffer nekukurumidza kana ikozvino yaiswa.
Equipment nokuda Polyacrylamide Gel Electrophoresis
Gel Electrophoresis Sero (Tangi/Chamber)
Iyo gel tangi ye polyacrylamide gel electrophoresis (PEJI) yakasiyana neagarose gel tank. Iyo agarose gel tangi yakachinjika, nepo PEJI tangi yakamira. Ne vertical electrophoresis cell (tangi/chamber), gel nhete (kazhinji 1.0mm kana 1.5mm) inodururwa pakati pemagirazi maviri egirazi uye inoiswa kuitira kuti pasi pejeri rinyudzwe mubuffer mune imwe kamuri uye kumusoro kunonyudzwa mubuffer. mune imwe kamuri. Kana yazvino ikaiswa, chitsama chidiki chebuffer chinotama kuburikidza nejeri kubva pakamuri yepamusoro kuenda kukamuri yepasi. Nezvibatiso zvakasimba zvekuvimbisa kuti gungano rinogara rakati twasu, mudziyo unofambisa girosi rinomhanya nekutonhodza zvichikonzera mabhandi akasiyana.
Beijing Liuyi Biotechnology Co., Ltd. (Liuyi Biotechnology) inogadzira huwandu hwehukuru hwe polyacrylamide gel electrophoresis masero (matangi/makamuri). Iwo mamodheru DYCZ-20C uye DYCZ-20G ari vertical electrophoresis maseru (matangi/makamuri) eDNA sequencing ongororo. Mamwe vertical electrophoresis maseru (matangi/makamuri) anoenderana ne blotting system, senge modhi DYCZ-24DN, DYCZ-25D uye DYCZ-25E inoenderana neWestern Blotting system modhi DYCZ-40D, DYCZ-40G uye DYCZ-40F, ayo anoshandiswa kuendesa mapuroteni molecule kubva mujeli kuenda ku membrane. Mushure meSDS-PAGE electrophoresis, Western Blotting inyanzvi yekuona puroteni chaiyo mumusanganiswa weprotein. Iwe unogona kusarudza idzi blotting masisitimu zvichienderana nekuyedza zvinodiwa.
Electrophoresis Power Supply
Kuti upe magetsi ekushandisa gel, iwe uchada electrophoresis magetsi. PaLiuyi Biotechnology tinopa huwandu hwemagetsi emagetsi emagetsi kune ese maapplication. Iyo modhi yeDYY-12 uye DYY-12C ine yakakwira yakagadzikana voltage uye yazvino inogona kusangana nepamusoro voltage inodiwa electrophoresis. Iyo ine basa rekumira, nguva, VH uye nhanho-ne-nhanho application. Iwo akanakira IEF uye DNA sequencing electrophoresis application. Kune zvakajairika mapuroteni uye DNA electrophoresis application, isu tine modhi yeDYY-2C, DYY-6C, DYY-10, zvichingodaro, izvo zvakare zvinopisa zvekutengesa magetsi magetsi ane electrophoresis maseru (matangi/makamuri). Izvi zvinogona kushandiswa kune yepakati uye yakaderera voltage electrophoresis application, senge yekushandisa lab yechikoro, lab yechipatara uye zvichingodaro. Mamwe mamodheru emagetsi ekushandisa, ndapota shanyira webhusaiti yedu.
Iyo Liuyi mhando ine inopfuura makore makumi mashanu nhoroondo muChina uye kambani inogona kupa yakagadzikana uye yemhando yepamusoro zvigadzirwa pasi rese. Kuburikidza nekukura kwemakore, yakakodzera sarudzo yako!
Kuti uwane rumwe ruzivo nezvedu, ndapota taura nesu neemail[email yakachengetedzwa] or [email yakachengetedzwa].
Mareferensi eChii chinonzi polyacrylamide gel electrophoresis?
1. Karen Steward PhD Polyacrylamide gel electrophoresis, Mashandiro Arinoita, Matanho akasiyana, uye mashandisirwo aro.
Nguva yekutumira: May-23-2022