Teko Molao-motheo
Hemoglobin electrophoresis e ikemiselitse ho lemoha le ho netefatsa li-hemoglobin tse fapaneng tse tloaelehileng le tse sa tloaelehang.
Ka lebaka la litefiso tse fapaneng le lintlha tsa isoelectric tsa mefuta e fapaneng ea hemoglobin, tharollong e itseng ea pH buffer, ha ntlha ea isoelectric ea hemoglobin e le tlase ho pH ea tharollo ea buffer, hemoglobin e jara tefiso e mpe ebe e fallela ho anode nakong ea electrophoresis. Ka lehlakoreng le leng, hemoglobin e nang le tefiso e ntle e fetela ho cathode.
Tlas'a motlakase o itseng le ka mor'a nako e itseng ea electrophoresis, li-hemoglobin tse nang le litefiso tse sa tšoaneng le boima ba limolek'hule li bontša litsela tse fapaneng tsa ho falla le lebelo. Sena se lumella karohano ea libaka tse ikhethileng, 'me tlhahlobo e latelang ea colorimetric kapa electrophoretic scanning e ka etsoa libakeng tsena ho lekanya hemoglobin e fapaneng. Mokhoa o sebelisoang haholo ke pH 8.6 cellulose acetate membrane electrophoresis.
Ka har'a cytoplasm, lihlopha tsa ethylene glycol (CHOH-CHOH) tse teng ka glycogen kapa polysaccharide lintho (tse kang mucopolysaccharides, mucoproteins, glycoproteins, glycolipids, joalo-joalo) li na le oxidized ke periodic acid 'me li fetoloa lihlopha tsa aldehyde (CHO-CHO). Lihlopha tsena tsa aldehyde li kopana le reagent ea Schiff e se nang 'mala e bopherese bo bofubelu, ho etsa dae e khubelu e pherese e behang moo li-polysaccharides li leng teng ka seleng. Boitšoaro bona bo tsejoa e le "periodic acid-Schiff" (PAS) staining, eo pele e neng e bitsoa glycogen staining.
Mokhoa oa ho Leka
Lisebelisoa:Cellulose acetatemembrane, lisebelisoa tsa electrophoresis(DYCP-38C le phepelo ea motlakase DYY-6C), Sesebelisoa sa Superior Sample Loading (pipette), spectrophotometer, colorimetric cuvettes, li-buffers.
Buffer:
(1) pH 8.6 TEB Buffer: Beha 10.29 g Tris, 0.6 g EDTA, 3.2 g boric acid, mme o kenye metsi a distilled ho 1000 ml.
(2) Borate Buffer: Beha 6.87 g borax le 5.56 g boric acid, 'me u kenye metsi a hloekisitsoeng ho 1000 ml.
Mokhoa:
Ptokiso ea Tharollo ea Hemoglobin
Nka 3 ml ea mali a nang le heparin kapa sodium citrate e le anticoagulant. Centrifuge ka 2000 rpm bakeng sa metsotso e 10 'me u lahle plasma. Hlatsoa lisele tse khubelu tsa mali ka makhetlo a mararo ka saline ea 'mele (750 rpm, metsotso e 5 centrifugation nako le nako). Centrifuge ka 2200 rpm bakeng sa metsotso e 10 'me u lahle matla a maholo. Eketsa palo e lekanang ea metsi a silafetseng, ebe u eketsa makhetlo a 0.5 bophahamo ba carbon tetrachloride. Shake ka matla bakeng sa metsotso ea 5, 'me joale centrifuge ka 2200 rpm bakeng sa metsotso ea 10 ho bokella tharollo e ka holimo ea Hb bakeng sa tšebeliso ea hamorao.
Ho kolobisa Membrane
Khaola lesela la cellulose acetate ka likhoele tse ka bang 3 cm × 8 cm. Li inele ka pH 8.6 TEB buffer ho fihlela li tletse ka botlalo, ebe u li tlosa 'me u li hlakole ka pampiri ea sefene.
Spotting
Sebelisa pipette ho bona 10 μl ea tharollo ea hemoglobin e otlolohileng holim'a lera la cellulose acetate (lehlakoreng le makukuno), le ka bang 1.5 cm ho tloha pheletsong.
Electrophoresis
Tšela tharollo ea borate buffer ka kamoreng ea electrophoresis. Beha lesela la cellulose acetate le lehlakoreng le nang le matheba qetellong ea cathode ea kamore. Matha ka 200 V bakeng sa metsotso e 30.
Elution
Khaola libaka tsa HbA le HbA2, u li behe ka har'a li-tubes tse arohaneng tsa teko, 'me u kenye 15 ml le 3 ml ea metsi a hloekisitsoeng, ka ho latellana. Ka bonolo sisinya ho elute haemoglobin ka ho feletseng, ka nako eo kopanya.
Colorimetry
Zero ho monya ho sebelisa metsi a hloekisitsoeng bakeng sa tharollo ea elution le ho lekanya ho monya ho 415 nm.
Palo
HbA2(%) = Ho se be le HbA2 tube / (Ho tlosoa ha HbA tube × 5 + Ho tlosoa ha HbA2 tube) × 100%
Palo ea Liphetho tsa Teko
Reference Range bakeng sa pH 8.6 TEB Buffer Cellulose Acetate Electrophoresis: HbA > 95%, HbA2 1% -3.1%
Lintlha
Nako ea electrophoresis ha ea lokela ho ba telele haholo. Lera la cellulose acetate ha lea lokela ho omella nakong ea electrophoresis. Emisa electrophoresis ha HbA le HbA2 li arohane ka ho hlaka. Electrophoresis ea nako e telele e ka baka ho hasana ha sehlopha le ho fifala.
Qoba ho sebelisa sampole e ngata haholo. Mokelikeli o mongata oa hemoglobin o ka lebisa ho khaohaneng ha lihlopha kapa litšila tse sa lekaneng, ho bakang maemo a phahameng a HbA.
Thibela tšilafalo ea lera la cellulose acetate ka liprotheine.
Mokelikeli ha oa lokela ho ba holimo haholo; ho seng joalo, lihlopha tsa hemoglobin li ka ’na tsa se ke tsa arohana.
Kamehla u kenyeletse mehlala ea batho ba tloaelehileng le li-hemoglobin tse hlokahalang tse tsejoang e le taolo.
Beijing Liuyi Biotechnology e etsa tanka ea profeshenale ea electrophoresis bakeng sa hemoglobin electrophoresis eo e leng mohlala.DYCP-38Ccellulose acetate membrane electrophoresis tank, mme ho na le mefuta e 'meli ea phepelo ea motlakase ea electrophoresis e fumanehang bakeng sa tanka ea cellulose acetate membrane electrophoresis.DYY-2CleDYY-6Cphepelo ea motlakase.
Ho sa le joalo, Beijing Liuyi Biotechnology e fana ka membrane ea cellulose acetate bakeng sa bareki, 'me boholo ba lesela la cellulose acetate bo ka etsoa ka mokhoa o ikhethileng. Rea u amohela ho re botsa lisampole le lintlha tse ling.
Letšoao la Beijing 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 lefats'eng ka bophara. Ho theosa le lilemo tsa nts'etsopele, e loketse khetho ea hau!
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Nako ea poso: Sep-20-2023