Ụkpụrụ nnwale
Hemoglobin electrophoresis na-achọ ịchọpụta ma gosipụta hemoglobin dị iche iche nkịtị na nke na-adịghị mma.
N'ihi ụgwọ dị iche iche na isi ihe isoelectric nke ụdị hemoglobin dị iche iche, na ụfọdụ pH buffer ngwọta, mgbe ebe dịpụrụ adịpụ nke hemoglobin dị ala karịa pH nke ngwọta nchekwa, hemoglobin na-ebu ụgwọ na-adịghị mma ma na-akwaga na anode n'oge electrophoresis. N'aka nke ọzọ, haemoglobin nwere ụgwọ dị mma na-aga n'ihu cathode.
N'okpuru ụfọdụ voltaji na mgbe oge electrophoresis a kapịrị ọnụ, hemoglobins nwere ụgwọ dị iche iche na nha molekụla na-egosiputa ntụzịaka na ọsọ ọsọ dị iche iche. Nke a na-enye ohere maka nkewa nke mpaghara dị iche iche, na-esote colorimetric ma ọ bụ electrophoretic scanning nwere ike ime na mpaghara ndị a iji chọpụta hemoglobin dị iche iche. Ụzọ a na-ejikarị eme ihe bụ pH 8.6 cellulose acetate membrane electrophoresis.
N'ime cytoplasm, otu ethylene glycol (CHOH-CHOH) dị na glycogen ma ọ bụ ihe polysaccharide (dị ka mucopolysaccharides, mucoproteins, glycoproteins, glycolipids, wdg) na-eme ka ọ bụrụ oxidized site na oge oge ma ghọọ otu aldehyde (CHO-CHO). Ndị otu aldehyde ndị a na-ejikọta ya na Schiff reagent na-acha ọbara ọbara na-acha ọbara ọbara, na-akpụ acha odo odo-acha ọbara ọbara nke na-edobe ebe polysaccharides dị na cell. A maara mmeghachi omume a dị ka ntụpọ acid-Schiff (PAS), nke a na-akpọbu glycogen ntụpọ.
Ụzọ nnwale
Akụrụngwa:Cellulose acetatemembrane, electrophoresis ngwa(DYCP-38C na ike ọkọnọ DDY-6C), Ngwá ọrụ nkwụnye ihe nlele kachasị elu(pipette), spectrophotometer, colorimetric cuvettes, buffers.
Nchekwa:
(1) pH 8.6 TEB Buffer: tụọ 10.29 g Tris, 0.6 g EDTA, 3.2 g boric acid, ma tinye mmiri na-esi ísì ụtọ na 1000 ml.
(2) Borate Buffer: Tụnyere 6.87 g borax na 5.56 g boric acid, ma tinye mmiri a na-ekpo ọkụ na 1000 ml.
Usoro:
Pmmezi nke hemoglobin Solution
Were 3 ml nke ọbara nwere heparin ma ọ bụ sodium citrate dị ka ọgwụ mgbochi. Centrifuge na 2000 rpm maka nkeji 10 wee tụfuo plasma. Jiri saline physiological sachaa mkpụrụ ndụ ọbara uhie ugboro atọ (750 rpm, centrifugation 5 nkeji oge ọ bụla). Centrifuge na 2200 rpm maka nkeji 10 wee tụfuo ihe dị elu. Gbakwunye mmiri a na-ekpochapụ nha nhata, wee gbakwunye okpukpu 0,5 nke carbon tetrachloride. Ma jijiji nke ukwuu maka nkeji 5, wee kee centrifuge na 2200 rpm maka nkeji 10 iji nakọta ihe ngwọta Hb dị elu maka iji mechaa.
Na-agbanye Membrane
Bee akpụkpọ ahụ cellulose acetate n'ime ibe nke 3 cm × 8 cm. Tinye ha na pH 8.6 TEB nchekwa ruo mgbe juputara n'uju, wee wepụ ma hichapụ ya na akwụkwọ nzacha.
Ntụpọ
Jiri pipette hụ 10 μl nke hemoglobin ngwọta kwụ ọtọ na cellulose acetate membrane (akụkụ siri ike), ihe dịka 1.5 cm site na nsọtụ.
Electrophoresis
Wunye ngwọta borate n'ime ụlọ electrophoresis. Debe akpụkpọ ahụ acetate cellulose na akụkụ ahụrụ anya na njedebe cathode nke ụlọ ahụ. Gbaa na 200 V maka nkeji 30.
Elution
Beepụ mpaghara HbA na HbA2, tinye ha n'ime akpa ule dị iche iche, tinye 15 ml na 3 ml nke mmiri a na-ekpo ọkụ n'otu n'otu. Jiri nwayọ maa jijiji ka o wee kpochapụ haemoglobin kpamkpam, wee gwakọta ya.
Colorimetry
Zero nnabata ahụ site na iji mmiri distilled maka ngwọta elution wee tụọ nnabata ahụ na 415 nm.
Mgbakọ
HbA2(%) = mmịpụta nke tube HbA2 / (Mmiri nke tube HbA2 × 5 + mmịpụta nke tube HbA2) × 100%
Mgbakọ nsonaazụ nnwale
Oke ntụaka maka pH 8.6 TEB Buffer Cellulose Acetate Electrophoresis: HbA> 95%, HbA2 1%-3.1%
Ihe ndetu
Oge Electrophoresis ekwesịghị ịdị ogologo. Akpụkpọ anụ acetate cellulose ekwesịghị ịkpọ nkụ n'oge electrophoresis. Kwụsị electrophoresis mgbe HbA na HbA2 kewapụrụ nke ọma. Electrophoresis na-adịte aka nwere ike ime ka mgbasa bandwit na blurring.
Zere iji ọtụtụ ihe nlele. Mmiri haemoglobin karịrị akarị nwere ike ibute mwepu eriri ma ọ bụ enweghị ntụpọ zuru oke, na-ebute ọkwa HbA n'ụzọ ụgha.
Gbochie mmetọ nke cellulose acetate membrane na protein.
Ihe dị ugbu a ekwesịghị ịdị elu; ma ọ bụghị ya, eriri hemoglobin nwere ike ọ gaghị ekewa.
Na-agụnye mgbe niile ụdịdị sitere na ndị nkịtị yana hemoglobins na-adịghị mma mara dị mkpa dị ka njikwa.
Beijing Liuyi Biotechnology na-arụpụta tankị electrophoresis ọkachamara maka hemoglobin electrophoresis bụ ihe atụ.DYCP-38Ccellulose acetate membrane electrophoresis tank, na e nwere ụdị abụọ nke electrophoresis ike ọkọnọ maka cellulose acetate membrane electrophoresis tank.DDY-2CnaMMỤỌ-6Cịnye ọkụ.
Ka ọ dị ugbu a, Beijing Liuyi Biotechnology na-enye akpụkpọ anụ acetate cellulose maka ndị ahịa, na nha cellulose acetate nwere ike ịhazi ya. Nabata ịjụ anyị maka nlele na ozi ndị ọzọ.
Akara Beijing Liuyi nwere ihe karịrị afọ 50 akụkọ ihe mere eme na China na ụlọ ọrụ ahụ nwere ike ịnye ngwaahịa kwụsiri ike na nke dị elu gburugburu ụwa. Site na afọ nke mmepe, ọ bụ ihe kwesịrị ekwesị nke nhọrọ gị!
Anyị na-achọ ugbu a ndị mmekọ, ma OEM electrophoresis tank na nkesa na-anabata.
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Oge nzipu: Sep-20-2023