{"id":3050,"date":"2016-05-22T17:28:52","date_gmt":"2016-05-22T16:28:52","guid":{"rendered":"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/?p=3050"},"modified":"2016-05-22T17:28:52","modified_gmt":"2016-05-22T16:28:52","slug":"western-blot","status":"publish","type":"post","link":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/modul-b\/western-blot\/","title":{"rendered":"Western Blot"},"content":{"rendered":"<p><a href=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_100728-2.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone wp-image-3055 size-large\" src=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_100728-2-1024x576.jpg\" alt=\"20160422_100728-2\" width=\"1024\" height=\"576\" srcset=\"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_100728-2-1024x576.jpg 1024w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_100728-2-300x169.jpg 300w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><\/a><\/p>\n<p>Com a estudiant del Grau Superior en Qu\u00edmica Ambiental estic realitzant les pr\u00e0ctiques de la FCT (Formaci\u00f3 en Centres de Treball) a l&#8217;<a href=\"http:\/\/www.idibell.cat\/en\" target=\"_blank\">Institut d\u2019Investigaci\u00f3 Biom\u00e8dica de Bellvitge (IDIBELL)<\/a>, concretament al <a href=\"http:\/\/www.idibell.cat\/modul\/biological-clues-of-the-invasive-and-metastasic-phenotype\/en\" target=\"_blank\">Laboratori d\u2019Oncologia Molecular<\/a>.<\/p>\n<p>Una de les t\u00e8cniques m\u00e9s utilitzada en l\u2019\u00e0mbit de la recerca biom\u00e8dica\u00a0\u00e9s el <a href=\"https:\/\/scykness.wordpress.com\/2013\/07\/16\/entendiendo-un-western-blot-o-al-menos-intentandolo\/\" target=\"_blank\">western blot<\/a> perqu\u00e8 ens permet identificar la concentraci\u00f3 d\u2019una determinada prote\u00efna en un medi com, per exemple, un cultiu cel\u00b7lular. A continuaci\u00f3, explicar\u00e9 de manera sintetitzada com es realitza i perqu\u00e8 serveix aquest m\u00e8tode i, d\u2019aquesta manera, podreu fer-vos una idea de com \u00e9s la meva experi\u00e8ncia i quines tasques desenvolupo al laboratori.<\/p>\n<p><!--more--><\/p>\n<p>El grup d\u2019investigaci\u00f3 del qual formo part es dedica a l\u2019estudi dels <a href=\"https:\/\/es.wikipedia.org\/wiki\/Hepatocarcinoma\" target=\"_blank\">hepatocarcinomes<\/a>, \u00e9s a dir, les c\u00e8l\u00b7lules canceroses del fetge. L&#8217;objectiu principal del grup \u00e9s veure com reaccionen aquestes c\u00e8l\u00b7lules\u00a0a la pres\u00e8ncia de diferents prote\u00efnes o reactius per tal de\u00a0desenvolupar nous tractaments.<\/p>\n<p><a href=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/Grup-Isabel-Fabregat-2016-2.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"  wp-image-3053 aligncenter\" src=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/Grup-Isabel-Fabregat-2016-2.jpg\" alt=\"Grup Isabel Fabregat 2016\" width=\"446\" height=\"335\" srcset=\"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/Grup-Isabel-Fabregat-2016-2.jpg 640w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/Grup-Isabel-Fabregat-2016-2-300x225.jpg 300w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/Grup-Isabel-Fabregat-2016-2-200x150.jpg 200w\" sizes=\"auto, (max-width: 446px) 100vw, 446px\" \/><\/a><\/p>\n<p>El m\u00e8tode del western blot ens permet veure com reacciona un cultiu cel\u00b7lular a unes determinades condicions, per exemple, ens permet saber si un enzim estimula o inhibeix creixement d\u2019un determinat cultiu cel\u00b7lular. Per veure com reacciona un cultiu cel\u00b7lular analitzarem la quantitat de prote\u00efna que cont\u00e9. Un cop explicat perqu\u00e8 s\u2019utilitza procedir\u00e9 a explicar el m\u00e8tode en s\u00ed. Conv\u00e9 destacar que l\u2019explicaci\u00f3 seg\u00fcent \u00e9s general i sintetitzada tenint en compte el grau de complexitat del m\u00e8tode.<\/p>\n<p>En primer lloc, agafarem les plaques de Petri que ens interessin i extraurem la prote\u00efna del cultiu utilitzant PBS i un <i>scraper<\/i>. Seguidament, recollirem la prote\u00efna en un tub d\u2019assaig i la centrifugarem per separar la prote\u00efna dels reactius utilitzats per extreure-la. Quan tinguem la prote\u00efna a\u00efllada la guardarem al congelador per conservar-la en condicions \u00f2ptimes.<\/p>\n<p>A continuaci\u00f3, procedirem a preparar un gel de poliacrilamida per realitzar una <a href=\"https:\/\/www.youtube.com\/watch?v=_8xftsCIwYo\" target=\"_blank\">electroforesi<\/a>. L\u2019electroforesi \u00e9s un m\u00e8tode de separaci\u00f3 de prote\u00efnes segons el seu pes molecular utilitzant c\u00e0rrega el\u00e8ctrica.\u00a0Per a preparar el gel que utilitzarem en l\u2019electroforesi necessitarem els seg\u00fcents reactius:<\/p>\n<ul>\n<li>Acrylamida<\/li>\n<li>SDS<\/li>\n<li>TEMED<\/li>\n<li>APS<\/li>\n<li>TRIS<\/li>\n<\/ul>\n<p><a href=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_095432-2.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"  wp-image-3071 aligncenter\" src=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_095432-2-1024x576.jpg\" alt=\"20160422_095432-2\" width=\"539\" height=\"303\" srcset=\"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_095432-2-1024x576.jpg 1024w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_095432-2-300x169.jpg 300w\" sizes=\"auto, (max-width: 539px) 100vw, 539px\" \/><\/a><\/p>\n<p>&nbsp;<\/p>\n<p>Un cop tenim el gel preparat ja podem realitzar l\u2019electroforesi. En aplicar un determinat voltatge a les prote\u00efnes, aquestes reaccionen de manera diferent segons el seu pes molecular i la seva concentraci\u00f3.<\/p>\n<p>Seguidament, transferirem les prote\u00efnes del gel de poliacrilamida a una membrana de nitrocel\u00b7lulosa.<\/p>\n<p>Per detectar una determinada prote\u00efna utilitzarem un antic\u00f2s primari i, de la mateixa manera, utilitzarem un antic\u00f2s secundari concret per detectar al primari en q\u00fcesti\u00f3. A continuaci\u00f3, posarem la membrana en contacte amb un reactiu anomenat ECL que fa que poguem visualitzar les prote\u00efnes com si es tract\u00e9s d&#8217;una\u00a0radiografia en forma de bandes.<\/p>\n<p>L\u2019\u00faltim pas \u00e9s el revelatge de les membranes. Introduirem les membranes en un <em>cassette<\/em> amb una pel\u00b7l\u00edcula fotogr\u00e0fica en la qual apareixeran bandes fosques com a resposta a l\u2019activitat radioactiva de la membrana. Les bandes fosques que apareixen a la pel\u00b7l\u00edcula fotogr\u00e0fica ens indiquen la concentraci\u00f3 d\u2019una determinada prote\u00efna en un cultiu cel\u00b7lular. El temps d&#8217;exposici\u00f3 variar\u00e0 segons els reactius i anticossos utilitzats.<\/p>\n<p><a href=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/MMR-07-05-1487-g06.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"  wp-image-3074 aligncenter\" src=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/MMR-07-05-1487-g06-902x1024.jpg\" alt=\"MMR-07-05-1487-g06\" width=\"362\" height=\"411\" srcset=\"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/MMR-07-05-1487-g06-902x1024.jpg 902w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/MMR-07-05-1487-g06-264x300.jpg 264w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/MMR-07-05-1487-g06.jpg 1087w\" sizes=\"auto, (max-width: 362px) 100vw, 362px\" \/><\/a><\/p>\n<p>&nbsp;<\/p>\n<p>La realitzaci\u00f3 d\u2019aquestes pr\u00e0ctiques ha estat una oportunitat \u00fanica per con\u00e8ixer de primera m\u00e0 el m\u00f3n de la investigaci\u00f3 biom\u00e8dica i estic segur de qu\u00e8 els coneixements que estic adquirint al llarg d\u2019aquests sis mesos em seran de gran utilitat en un futur pr\u00f2xim a l\u2019hora d\u2019entrar al mercat laboral. A m\u00e9s, per a mi \u00e9s tot un plaer aprendre de professionals del m\u00f3n de la recerca\u00a0com ho s\u00f3n els meus companys de laboratori i els agraeixo profundament l\u2019amabilitat i paci\u00e8ncia que estan tenint amb mi, gr\u00e0cies a ells el meu aprenentatge i formaci\u00f3 creixen\u00a0exponencialment dia a dia.<\/p>\n<p><a href=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_103045-2.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"  wp-image-3079 aligncenter\" src=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_103045-2-1024x576.jpg\" alt=\"20160422_103045-2\" width=\"605\" height=\"340\" srcset=\"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_103045-2-1024x576.jpg 1024w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_103045-2-300x169.jpg 300w\" sizes=\"auto, (max-width: 605px) 100vw, 605px\" \/><\/a><\/p>\n<p><a href=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_093017-3-2.jpg\"><img loading=\"lazy\" decoding=\"async\" class=\"  wp-image-3078 aligncenter\" src=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_093017-3-2-1024x576.jpg\" alt=\"20160422_093017-3-2\" width=\"615\" height=\"346\" srcset=\"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_093017-3-2-1024x576.jpg 1024w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/20160422_093017-3-2-300x169.jpg 300w\" sizes=\"auto, (max-width: 615px) 100vw, 615px\" \/><\/a><\/p>\n<p>&nbsp;<\/p>\n<p><a href=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/SDS-PAGE_Electrophoresis.png\"><img loading=\"lazy\" decoding=\"async\" class=\"alignnone size-full wp-image-3081\" src=\"http:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/SDS-PAGE_Electrophoresis.png\" alt=\"SDS-PAGE_Electrophoresis\" width=\"800\" height=\"275\" srcset=\"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/SDS-PAGE_Electrophoresis.png 800w, https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-content\/uploads\/usu644\/2016\/05\/SDS-PAGE_Electrophoresis-300x103.png 300w\" sizes=\"auto, (max-width: 800px) 100vw, 800px\" \/><\/a><\/p>\n<p>&nbsp;<\/p>\n<div class=\"ast-oembed-container \" style=\"height: 100%;\"><iframe loading=\"lazy\" title=\"Western Blotting\" width=\"500\" height=\"281\" src=\"https:\/\/www.youtube.com\/embed\/VgAuZ6dBOfs?feature=oembed\" frameborder=\"0\" allow=\"accelerometer; autoplay; clipboard-write; encrypted-media; gyroscope; picture-in-picture; web-share\" referrerpolicy=\"strict-origin-when-cross-origin\" allowfullscreen><\/iframe><\/div>\n","protected":false},"excerpt":{"rendered":"<p>Com a estudiant del Grau Superior en Qu\u00edmica Ambiental estic realitzant les pr\u00e0ctiques de la FCT (Formaci\u00f3 en Centres de [&hellip;]<\/p>\n","protected":false},"author":55,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","theme-transparent-header-meta":"","adv-header-id-meta":"","stick-header-meta":"","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"var(--ast-global-color-4)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"footnotes":""},"categories":[268,174],"tags":[291,293,290,292,289],"class_list":["post-3050","post","type-post","status-publish","format-standard","hentry","category-fct","category-modul-b","tag-biomedicina","tag-electroforesi","tag-investigacio","tag-laboratori","tag-western-blot"],"post_mailing_queue_ids":[],"_links":{"self":[{"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/posts\/3050","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/users\/55"}],"replies":[{"embeddable":true,"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/comments?post=3050"}],"version-history":[{"count":3,"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/posts\/3050\/revisions"}],"predecessor-version":[{"id":3090,"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/posts\/3050\/revisions\/3090"}],"wp:attachment":[{"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/media?parent=3050"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/categories?post=3050"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/agora.xtec.cat\/ies-merce-rodoreda\/wp-json\/wp\/v2\/tags?post=3050"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}