He aha ka polyacrylamide gel electrophoresis?

Polyacrylamide Gel Electrophoresis

ʻO ka electrophoresis gel kahi ʻenehana kumu i loko o nā laboratories ma waena o nā hoʻopaʻapaʻa biological, e ʻae i ka hoʻokaʻawale ʻana o nā macromolecules e like me DNA, RNA a me nā protein. Hiki i nā ʻenehana hoʻokaʻawale ʻokoʻa a me nā mīkini ke hoʻokaʻawale maikaʻi i nā ʻāpana o kēia mau molekala ma o ka hoʻohana ʻana i ko lākou ʻano kino. No nā protein, ʻo ka polyacrylamide gel electrophoresis (PAGE) ke ʻano o ke koho.

ʻO ka PAGE kahi hana e hoʻokaʻawale ai i nā macromolecules e like me nā protein e pili ana i kā lākou electrophoretic mobility, ʻo ia hoʻi, ka hiki o nā analytes ke neʻe i kahi electrode o ka uku kū'ē. I ka PAGE, hoʻoholo ʻia kēia e ka uku, ka nui (ke kaumaha mole) a me ke ʻano o ka mole. Ke neʻe nei nā analytes ma nā pores i hana ʻia i ka gel polyacrylamide. ʻAʻole like me DNA a me RNA, ʻokoʻa nā protein e like me nā amino acids i hoʻohui ʻia, hiki ke hoʻololi i ka holo ʻana. Hiki i nā kaula amino acid ke hana i nā hale kiʻekiʻe e pili ana i ko lākou nui a no laila pehea e hiki ai iā lākou ke neʻe i loko o nā pores. No laila, makemake ʻia i kekahi manawa e hoʻopau i nā protein ma mua o ka electrophoresis e hoʻokaʻina iā lākou inā makemake ʻia kahi kuhi pololei o ka nui.

SDS PAGE

ʻO ka sodium-dodecyl sulfate polyacrylamide gel electrophoresis kahi ʻenehana i hoʻohana ʻia e hoʻokaʻawale i nā molekala protein o ka nui o 5 a 250 kDa. Hoʻokaʻawale ʻia nā protein ma muli o ko lākou kaumaha molekala. Hoʻohui ʻia ʻo Sodium dodecyl sulfate, kahi surfactant anionic, i ka hoʻomākaukau ʻana i nā gels e uhi ana i nā koina koʻikoʻi o nā ʻāpana protein a hāʻawi iā lākou i ka uku like me ka ratio nuipa. Ma nā huaʻōlelo maʻalahi, hoʻokaʻawale ia i nā protein a hāʻawi iā lākou i ka uku maikaʻi ʻole.

ʻAO ʻŌiwi

ʻO ka PAGE maoli kahi hana e hoʻohana ai i nā gels non-denatured no ka hoʻokaʻawale ʻana i nā protein. ʻAʻole like me SDS PAGE, ʻaʻohe mea denaturing i hoʻohui ʻia i ka hoʻomākaukau ʻana i nā gels. ʻO ka hopena, e hoʻokaʻawale ʻia nā protein ma ke kumu o ka uku a me ka nui o nā protein. Ma kēia ʻenehana, ʻo ka conformation, ka pelu ʻana a me nā kaulahao amino acid o nā protein nā kumu e hilinaʻi ai ka kaʻawale. ʻAʻole pōʻino nā protein i kēia kaʻina hana, a hiki ke hoʻihoʻi ʻia ma hope o ka pau ʻana o ka kaʻawale.

Pehea e hana ai ka polyacrylamide gel electrophoresis (PAGE)?

ʻO ke kumu kumu o ka PAGE ʻo ka hoʻokaʻawale ʻana i nā analytes ma ka hoʻohele ʻana iā lākou ma nā pores o kahi gel polyacrylamide me ka hoʻohana ʻana i ka uila. No ka hoʻokō ʻana i kēia, hoʻohui ʻia kahi hui acrylamide-bisacrylamide i polymerized (polyacrylamide) ma o ka hoʻohui ʻana i ka ammonium persulfate (APS). ʻO ka hopena, i hoʻopaʻa ʻia e ka tetramethylethylenediamine (TEMED), hana i kahi ʻupena e like me ka hale me nā pores e hiki ai i nā analytes ke neʻe (Figure 2). ʻO ke kiʻekiʻe o ka pākēneka o ka acrylamide a pau i hoʻokomo ʻia i loko o ka gel, ʻoi aku ka liʻiliʻi o ka nui o ka pore, no laila ʻoi aku ka liʻiliʻi o nā protein e hiki ke hele. ʻO ka ratio o ka acrylamide a hiki i ka bisacrylamide e pili pū i ka nui o ka pore akā mālama mau ʻia kēia. ʻO ka nui o nā pore liʻiliʻi e hoʻemi i ka wikiwiki e hiki ai i nā protein liʻiliʻi ke neʻe i loko o ka gel, e hoʻomaikaʻi ana i kā lākou hoʻonā a pale iā lākou mai ka holo wikiwiki ʻana i loko o ka buffer ke hoʻohana ʻia.

Mea lako no ka Polyacrylamide Gel Electrophoresis

Kelepona Electrophoresis Gel (Kahu/Keʻena)
ʻOkoʻa ka pahu gel no ka polyacrylamide gel electrophoresis (PAGE) mai ka pahu gel agarose. He paepae ka pahu gel agarose, a ʻo ka pahu PAGE ke kū pololei. Ma o ke kelepona electrophoresis vertical (tank/chamber), e ninini ʻia kahi gel lahilahi (maʻamau 1.0mm a i ʻole 1.5mm) ma waena o nā papa aniani ʻelua a kau ʻia i lalo o ka gel i loko o ka buffer i loko o hoʻokahi keʻena a hoʻopaʻa ʻia ka luna i loko o ka pahu. ma kekahi keena. Ke hoʻohana ʻia ka ʻāwīwī, neʻe ka nui o ka buffer ma o ka gel mai ke keʻena luna a i ke keʻena lalo. Me nā puʻupuʻu ikaika e hōʻoiaʻiʻo i ka noho ʻana o ka hui ma kahi kūlana kūpono, hoʻomaʻamaʻa nā mea hana i ka holo wikiwiki ʻana o ka gel me ka hoʻomaha ʻana i ka hopena i nā ʻāpana ʻokoʻa.

Ke hana nei ʻo Beijing Liuyi Biotechnology Co., Ltd. (Liuyi Biotechnology) i nā ʻano nui o ka polyacrylamide gel electrophoresis cell (tanks/chambers). ʻO nā hiʻohiʻona DYCZ-20C a me DYCZ-20G he mau pūnaewele electrophoresis vertical (nā pahu / keʻena) no ka nānā ʻana i ke kaʻina hana DNA. ʻO kekahi o nā pūnaewele electrophoresis kūpaʻa (nā pahu/keʻena) i kūpono me ka ʻōnaehana blotting, e like me ke kumu hoʻohālike DYCZ-24DN, DYCZ-25D a me DYCZ-25E kūpono me Western Blotting system model DYCZ-40D, DYCZ-40G a me DYCZ-40F, i hoʻohana ʻia no ka hoʻololi ʻana i ka molekele protein mai ka gel i ka membrane. Ma hope o ka SDS-PAGE electrophoresis, Western Blotting kahi hana e ʻike ai i kahi protein kikoʻī i loko o kahi hui pūmua. Hiki iā ʻoe ke koho i kēia mau ʻōnaehana blotting e like me nā koi hoʻokolohua.

Electrophoresis mana lako
No ka hāʻawi ʻana i ka uila no ka holo ʻana i ka gel, pono ʻoe i kahi mana electrophoresis. Ma Liuyi Biotechnology ke hāʻawi nei mākou i nā lako mana electrophoresis no nā noi āpau. Hiki i ke kŘkohu DYY-12 a me DYY-12C me ka uila pa'a ki'eki'e a me ka manawa e hiki ke ho'okō i ka electrophoresis koi. Loaʻa iā ia ka hana o ke kū, ka manawa, VH a me ka noi ʻana i kēlā me kēia. He kūpono lākou no ka noi electrophoresis IEF a me DNA. No ka protein maʻamau a me ka DNA electrophoresis noi, loaʻa iā mākou ke kumu hoʻohālike DYY-2C, DYY-6C, DYY-10, a pēlā aku, ʻo ia hoʻi nā lako mana kūʻai wela me nā cell electrophoresis (tanks/chambers). Hiki ke hoʻohana ʻia kēia mau mea no nā noi electrophoresis waena a me ka haʻahaʻa haʻahaʻa, e like me ka hoʻohana ʻana i ka lab kula, ke keʻena haukapila a pēlā aku. Nā hiʻohiʻona hou aku no nā lako mana, e ʻoluʻolu e kipa i kā mākou pūnaewele.

Loaʻa i ka brand Liuyi ma mua o 50 mau makahiki moʻolelo ma Kina a hiki i ka hui ke hāʻawi i nā huahana kūpaʻa a kiʻekiʻe a puni ka honua. Ma o ka ulu ʻana o nā makahiki, kūpono ia i kāu koho!

No ka ʻike hou aku e pili ana iā mākou, e ʻoluʻolu e kelepona mai iā mākou ma ka leka uila[palapala leka uila] or [palapala leka uila].

He aha ka polyacrylamide gel electrophoresis?
1. Karen Steward PhD Polyacrylamide gel electrophoresis, Pehea e hana ai, nā ʻano ʻenehana, a me kāna mau noi.