Polyacrylamide gel electrophoresis ke eng?

Polyacrylamide Gel Electrophoresis

Gel electrophoresis ke mokhoa oa mantlha lilaboratoring ho pholletsa le lithuto tsa baeloji, o lumellang karohano ea li-macromolecules tse kang DNA, RNA le liprotheine. Mecha ea litaba le mekhoa e fapaneng ea karohano e lumella li-subsets tsa limolek'hule tsena hore li arohane ka katleho ka ho sebelisa litšobotsi tsa tsona tsa 'mele. Bakeng sa liprotheine ka ho khetheha, polyacrylamide gel electrophoresis (PAGE) hangata ke mokhoa oa khetho.

LEQEPHE ke mokhoa o arolang li-macromolecules tse kang liprotheine tse thehiloeng ho motsamao oa tsona oa electrophoretic, ke hore, bokhoni ba li-analyte ho ea ho electrode ea tefiso e fapaneng. Ho LEQEPHE, sena se khethoa ke tefiso, boholo (boima ba molek'hule) le sebōpeho sa molek'hule. Bahlahlobisisi ba tsamaea ka li-pores tse entsoeng ka gel ea polyacrylamide. Ho fapana le DNA le RNA, liprotheine li fapana ka tefiso ho latela li-amino acid tse kenyellelitsoeng, tse ka susumetsang tsela eo li sebetsang ka eona. Likhoele tsa amino acid le tsona li ka etsa likarolo tsa bobeli tse amang boholo ba tsona, ka lebaka leo li khona ho tsamaea ka har'a masoba. Ka hona, ka linako tse ling ho ka ba ho lakatsehang ho hlahisa liprotheine pele ho electrophoresis ho li etsa linearize haeba ho hlokahala khakanyo e nepahetseng haholoanyane ea boholo.

SDS LEQEPHE

Sodium-dodecyl sulfate polyacrylamide gel electrophoresis ke mokhoa o sebelisoang ho arola limolek'hule tsa protheine tsa bongata ba 5 ho isa ho 250 kDa. Liprotheine li arotsoe feela ho latela boima ba tsona ba limolek'hule. Sodium dodecyl sulfate, e leng anionic surfactant, e eketsoa ho lokisoeng ha li-gel tse patang litefiso tsa lisampole tsa protheine le ho li fa tefiso e ts'oanang le tekanyo ea boima. Ka mantsoe a bonolo, e senya liprotheine mme e li fa tefiso e mpe.

Native LEQEPHE

Native PAGE ke mokhoa o sebelisang li-gel tse se nang denatured bakeng sa karohano ea liprotheine. Ho fapana le SDS PAGE, ha ho na moemeli oa denaturing o eketsoang ho lokisoeng ha li-gel. Ka lebaka leo, ho arohana ha liprotheine ho etsahala motheong oa tefiso le boholo ba liprotheine. Mohlaleng ona, liketane tsa liprotheine tsa conformation, mene le amino acid ke lintlha tseo karohano e itšetlehileng ka tsona. Liprotheine ha li senyehe ts'ebetsong ena, 'me li ka khutlisoa ka mor'a hore karohano e phethoe.

Polyacrylamide gel electrophoresis (PAGE) e sebetsa joang?

Molao-motheo oa motheo oa LEQEPHE ke ho arola bahlahlobisisi ka ho ba fetisa ka li-pores tsa gel ea polyacrylamide ho sebelisa motlakase oa motlakase. Ho finyella sena, motsoako oa acrylamide–bisacrylamide o etsoa polymerized (polyacrylamide) ka ho eketsoa ka ammonium persulfate (APS). Karabelo, e hlahisoang ke tetramethylethylenediamine (TEMED), e etsa letlooa le kang sebopeho se nang le li-pores tseo li-analyte li ka tsamaeang ka tsona (Setšoantšo sa 2). Ha karolo ea kakaretso ea acrylamide e kenyellelitsoeng ka gel e phahame, e fokotsa boholo ba pore, kahoo liprotheine tse tla khona ho feta li tla fokotseha. Karolelano ea acrylamide ho bisacrylamide le eona e tla ama boholo ba pore empa hangata sena se lula se le teng. Li-pore tse nyane li boetse li fokotsa lebelo leo liprotheine tse nyane li khonang ho tsamaea ka har'a gel, ho ntlafatsa tharollo ea tsona le ho li thibela ho matha ka har'a buffer ka potlako ha ho sebelisoa hona joale.

Thepa bakeng sa Polyacrylamide Gel Electrophoresis

Seleng ea Gel Electrophoresis (Tanka/Kamore)
Tanka ea gel bakeng sa polyacrylamide gel electrophoresis (PAGE) e fapane le tanka ea gel ea agarose. Tanka ea gel ea agarose e tšekaletse, ha tanka ea PAGE e otlolohile. Ka sele e otlolohileng ea electrophoresis (tanka / kamore), gel e tšesaane (ka tloaelo 1.0mm kapa 1.5mm) e tšeloa pakeng tsa lipoleiti tse peli tsa khalase ebe e kenngoa e le hore karolo e ka tlase ea gel e kenelle ka har'a buffer kamoreng e le 'ngoe' me e ka holimo e qoetsoe ka har'a buffer. phaposing enngwe. Ha motlakase o sebelisoa, palo e nyane ea buffer e tsamaea ka gel ho tloha phaposing e kaholimo ho ea phaposing e ka tlase. Ka li-clamp tse matla ho netefatsa hore kopano e lula e otlolohile, sesebelisoa se thusa hore gel e tsamaee ka potlako e nang le pholile e etsang hore ho be le lihlopha tse ikhethang.

Beijing Liuyi Biotechnology Co., Ltd. (Liuyi Biotechnology) e etsa mefuta e mengata ea boholo ba lisele tsa polyacrylamide gel electrophoresis (litanka / likamore). Mefuta ea DYCZ-20C le DYCZ-20G ke lisele tsa electrophoresis tse otlolohileng (litanka / likamore) bakeng sa tlhahlobo ea tatellano ea DNA. Tse ling tsa lisele tse otlolohileng tsa electrophoresis (litanka / likamore) li tsamaisana le sistimi ea ho koala, joalo ka mohlala oa DYCZ-24DN, DYCZ-25D le DYCZ-25E li tsamaisana le mohlala oa Western Blotting system DYCZ-40D, DYCZ-40G le DYCZ-40F, tse sebedisoang bakeng sa ho fetisetsa molek'hule wa protheine ho tloha gel ho ya ho lera. Ka mor'a SDS-PAGE electrophoresis, Western Blotting ke mokhoa oa ho lemoha protheine e itseng ka motsoako oa protheine. U ka khetha litsamaiso tsena tsa blotting ho latela litlhoko tsa liteko.

Phepelo ea Matla a Electrophoresis
Ho fana ka motlakase bakeng sa ho tsamaisa gel, o tla hloka motlakase oa electrophoresis. Ho Liuyi Biotechnology re fana ka mefuta e mengata ea lisebelisoa tsa motlakase tsa electrophoresis bakeng sa lits'ebetso tsohle. Mofuta oa DYY-12 le DYY-12C o nang le motlakase o tsitsitseng o phahameng le oa hona joale o ka kopana le tlhokahalo e phahameng ea motlakase oa electrophoresis. E na le ts'ebetso ea ho ema, nako, VH le ts'ebeliso ea mohato ka mohato. Li loketse IEF le DNA sequencing electrophoresis application. Bakeng sa ts'ebeliso e akaretsang ea protheine le DNA electrophoresis, re na le mohlala oa DYY-2C, DYY-6C, DYY-10, joalo-joalo, e leng lisebelisoa tsa motlakase tse chesang tse nang le lisele tsa electrophoresis (tanka / likamore). Tsena li ka sebelisoa bakeng sa lits'ebetso tsa electrophoresis tse mahareng le tse tlase, joalo ka ts'ebeliso ea lab ea sekolo, lab ea sepetlele joalo-joalo. Mefuta e meng ea lisebelisoa tsa motlakase, ka kopo etela sebaka sa rona sa marang-rang.

Letšoao la Liuyi le na le nalane ea lilemo tse fetang 50 Chaena mme k'hamphani e ka fana ka lihlahisoa tse tsitsitseng le tsa boleng bo holimo lefatšeng ka bophara. Ka nts'etsopele ea lilemo, e loketse khetho ea hau!

Ho fumana lintlha tse ling ka rona, ka kopo ikopanye le rona ka lengolo-tsoibila[imeile e sirelelitsoeng] or [imeile e sirelelitsoeng].

References for Polyacrylamide gel electrophoresis ke eng?
1. Karen Steward PhD Polyacrylamide gel electrophoresis, Kamoo e Sebetsang kateng, Mefuta e sa tšoaneng ea Mekhoa, le Tšebeliso ea eona