Kodi polyacrylamide gel electrophoresis ndi chiyani?

Polyacrylamide Gel Electrophoresis

Gel electrophoresis ndi njira yofunikira m'ma laboratories m'magulu onse achilengedwe, kulola kulekanitsa ma macromolecules monga DNA, RNA ndi mapuloteni. Zofalitsa zolekanitsa ndi njira zosiyanasiyana zimalola magawo a mamolekyuwa kuti apatulidwe bwino kwambiri pogwiritsa ntchito mawonekedwe awo. Kwa mapuloteni makamaka, polyacrylamide gel electrophoresis (PAGE) nthawi zambiri ndiyo njira yosankha.

PAGE ndi njira yomwe imalekanitsa ma macromolecules monga mapuloteni kutengera ma electrophoretic mobility, ndiko kuti, kuthekera kwa analytes kupita ku electrode yotsutsana nayo. Mu PAGE, izi zimatsimikiziridwa ndi mtengo, kukula (kulemera kwa molekyulu) ndi mawonekedwe a molekyulu. Ma analytes amadutsa pores opangidwa mu polyacrylamide gel. Mosiyana ndi DNA ndi RNA, mapuloteni amasiyana malinga ndi ma amino acid ophatikizidwa, omwe amatha kukhudza momwe amayendera. Zingwe za amino acid zimathanso kupanga zida zachiwiri zomwe zimakhudza kukula kwake komanso momwe zimadumphira m'mabowo. Chifukwa chake nthawi zina zingakhale zofunikila kupangira mapuloteni asanakhale ndi electrophoresis kuti azitha kuwatsata ngati kuyerekeza kolondola kwa kukula kumafunikira.

SDS PAGE

Sodium-dodecyl sulfate polyacrylamide gel electrophoresis ndi njira yomwe imagwiritsidwa ntchito kulekanitsa mamolekyu a mapuloteni amtundu wa 5 mpaka 250 kDa. Mapuloteni amasiyanitsidwa potengera kulemera kwawo kwa maselo. Sodium dodecyl sulfate, anionic surfactant, amawonjezedwa pokonza ma gels omwe amaphimba machulukidwe azinthu zama protein ndikuwapatsa mtengo wofanana ndi kuchuluka kwa misa. M'mawu osavuta, imasokoneza mapuloteni ndikuwapatsa chiwongola dzanja choyipa.

Nawo PAGE

Native PAGE ndi njira yomwe imagwiritsa ntchito ma gels osasinthika pakulekanitsa mapuloteni. Mosiyana ndi SDS PAGE, palibe wothandizira denaturing amawonjezeredwa pokonzekera ma gels. Chotsatira chake, kulekanitsa kwa mapuloteni kumachitika pamaziko a malipiro ndi kukula kwa mapuloteni. Munjira iyi, unyolo wa conformation, kupindika ndi amino acid wa mapuloteni ndizomwe zimapangitsa kulekanitsidwa kumadalira. Mapuloteni sakuwonongeka mwanjira iyi, ndipo amatha kubwezeredwa pambuyo pomaliza kulekanitsa.

Kodi polyacrylamide gel electrophoresis (PAGE) imagwira ntchito bwanji?

Mfundo yofunikira ya PAGE ndikulekanitsa owerengera powadutsa ma pores a gel osakaniza a polyacrylamide pogwiritsa ntchito magetsi. Kuti izi zitheke, kusakaniza kwa acrylamide–bisacrylamide kumapangidwa polima (polyacrylamide) powonjezera ammonium persulfate (APS). Zomwe zimapangidwira, zomwe zimapangidwira ndi tetramethylethylenediamine (TEMED), zimapanga ukonde wofanana ndi ma pores omwe ma analyte amatha kusuntha (Chithunzi 2). Kukwera kwa kuchuluka kwa acrylamide yophatikizidwa mu gel, kumapangitsa kuti pore ikhale yaying'ono, motero mapuloteni omwe amatha kudutsamo amakhala ochepa. Chiyerekezo cha acrylamide ku bisacrylamide chidzakhudzanso kukula kwa pore koma izi nthawi zambiri zimakhala zosasintha. Miyezo yaying'ono ya pore imachepetsanso liwiro lomwe mapuloteni ang'onoang'ono amatha kudutsa mu gel, kuwongolera mawonekedwe awo ndikulepheretsa kuti asathamangire ku buffer mwachangu akagwiritsidwa ntchito.

Zida za Polyacrylamide Gel Electrophoresis

Gel Electrophoresis Cell (Thanki/Chipinda)
Tanki ya gel ya polyacrylamide gel electrophoresis (PAGE) ndi yosiyana ndi thanki ya gel ya agarose. Tanki ya agarose gel ndi yopingasa, pomwe thanki ya PAGE imakhala yoyima. Ndi vertical electrophoresis cell (thanki/chipinda), gel osakaniza (kawirikawiri 1.0mm kapena 1.5mm) amatsanuliridwa pakati pa mbale ziwiri zamagalasi ndikuyikidwa kuti pansi pa gel osakaniza amizidwe mu chipinda chimodzi ndipo pamwamba pake amamizidwa mu buffer. m’chipinda china. Ikagwiritsidwa ntchito, chotchingira pang'ono chimasuntha kudzera mu gel kuchokera kuchipinda chapamwamba kupita kuchipinda chapansi. Ndi zingwe zolimba kuti zitsimikizire kuti msonkhanowo ukhala wowongoka, chipangizochi chimathandizira kuthamanga kwa gel osakaniza ndi ngakhale kuziziritsa komwe kumabweretsa mabandi apadera.

Beijing Liuyi Biotechnology Co., Ltd. (Liuyi Biotechnology) imapanga mitundu ingapo yama cell a polyacrylamide gel electrophoresis (matangi/zipinda). Mitundu ya DYCZ-20C ndi DYCZ-20G ndi ma cell electrophoresis (matangi / zipinda) zowunikira ma DNA. Ena ofukula ma cell electrophoresis (akasinja / zipinda) n'zogwirizana ndi blotting dongosolo, monga chitsanzo DYCZ-24DN, DYCZ-25D ndi DYCZ-25E n'zogwirizana ndi Western Blotting dongosolo chitsanzo DYCZ-40D, DYCZ-40G ndi DYCZ-40F, zomwe zimagwiritsidwa ntchito kusamutsa molekyulu ya protein kuchokera ku gel kupita ku nembanemba. Pambuyo pa SDS-PAGE electrophoresis, Western Blotting ndi njira yodziwira puloteni inayake mumsanganizo wa mapuloteni. Mutha kusankha makina opukutira awa molingana ndi zoyeserera.

Electrophoresis Power Supply
Kuti mupereke magetsi ogwiritsira ntchito gel osakaniza, mudzafunika magetsi a electrophoresis. Ku Liuyi Biotechnology timapereka magetsi osiyanasiyana a electrophoresis pazogwiritsa ntchito zonse. Mtundu wa DYY-12 ndi DYY-12C wokhala ndi magetsi okhazikika komanso apano amatha kukumana ndi ma electrophoresis ofunikira kwambiri. Ili ndi ntchito yoyimilira, nthawi, VH ndikugwiritsa ntchito pang'onopang'ono. Ndi abwino kwa IEF ndi DNA sequencing electrophoresis application. Pazambiri zama protein ndi DNA electrophoresis application, tili ndi mtundu wa DYY-2C, DYY-6C, DYY-10, ndi zina zotero, zomwe zilinso ndi mphamvu zogulitsa zotentha zokhala ndi ma cell a electrophoresis (matangi / zipinda). Izi zitha kugwiritsidwa ntchito pamagetsi apakati ndi otsika a electrophoresis, monga kugwiritsa ntchito labu yakusukulu, labu yakuchipatala ndi zina zotero. Mitundu yambiri yamagetsi, chonde pitani patsamba lathu.

Mtundu wa Liuyi uli ndi mbiri yopitilira zaka 50 ku China ndipo kampaniyo imatha kupereka zinthu zokhazikika komanso zapamwamba padziko lonse lapansi. Kupyolera mu chitukuko cha zaka, ndizoyenera kusankha kwanu!

Kuti mudziwe zambiri za ife, chonde titumizireni imelo[imelo yotetezedwa] or [imelo yotetezedwa].

Zolozera za Kodi polyacrylamide gel electrophoresis ndi chiyani?
1. Karen Steward PhD Polyacrylamide gel electrophoresis, Momwe Imagwirira Ntchito, Njira Zosiyanasiyana, ndi Kagwiritsidwe Ntchito Kake