{"id":1790,"date":"2021-09-16T14:36:11","date_gmt":"2021-09-16T14:36:11","guid":{"rendered":"https:\/\/lab.research.sickkids.ca\/sparc-molecular-analysis\/?page_id=1790"},"modified":"2021-09-16T14:36:11","modified_gmt":"2021-09-16T14:36:11","slug":"acetone-precipitation","status":"publish","type":"page","link":"https:\/\/lab.research.sickkids.ca\/sparc-molecular-analysis\/acetone-precipitation\/","title":{"rendered":"Acetone Precipitation"},"content":{"rendered":"<h1 style=\"text-align: center\"><span style=\"color: #000000\"><strong>PROTOCOL <\/strong><\/span><\/h1>\n<h3 style=\"text-align: center\"><span style=\"color: #000000\"><strong>ACETONE PRECIPITATION OF PROTEINS<\/strong><\/span><\/h3>\n<p><span style=\"color: #000000\"><em>This protocol can be used to precipitate proteins from solutions where heavy detergents were used to prepare protein extracts.<\/em> <\/span><\/p>\n<p><span style=\"color: #000000\"><strong>Required materials: <\/strong><\/span><\/p>\n<ul>\n<li><span style=\"color: #000000\">Cold (-20\u00b0C) acetone (volume = 4X that of the protein samples to be precipitated).<\/span><\/li>\n<li><span style=\"color: #000000\">Centrifuge tubes made of acetone-compatible polypropylene and able to hold 5X sample volume.<\/span><\/li>\n<li><span style=\"color: #000000\">Centrifuge and rotor for the tubes used (min speed 18000 x g).<\/span><\/li>\n<li><span style=\"color: #000000\">1M Urea\/50 mM NH4HCO3.<\/span><\/li>\n<\/ul>\n<p><span style=\"color: #000000\"><strong>Procedure:<\/strong><\/span><\/p>\n<ol>\n<li><span style=\"color: #000000\">Cool required volume of acetone to -20\u00b0C.<\/span><\/li>\n<li><span style=\"color: #000000\">Place protein sample into acetone-compatible tube.<\/span><\/li>\n<li><span style=\"color: #000000\">Add 4 fold volume of cold acetone to protein mixture.<\/span><\/li>\n<li><span style=\"color: #000000\">Vortex and incubate at -80\u00b0C for 1 hour (or overnight)<\/span><\/li>\n<li><span style=\"color: #000000\">Centrifuge @ 18,000g for 10 minutes<\/span><\/li>\n<li><span style=\"color: #000000\">Decant and properly dispose of supernatant, keeping in mind to not disturb the protein pellet.<\/span><\/li>\n<li><span style=\"color: #000000\">Allow pellet to air dry for 10-30 minutes in uncapped tube. Do not over-dry the pellet as it may not re-dissolve properly.<\/span><\/li>\n<li><span style=\"color: #000000\">Re-suspend the pellet in 50-100\u03bcL 1M Urea\/50mM NH4HCO3.<\/span><\/li>\n<li><span style=\"color: #000000\">Bring sample to SPARC facility OR, proceed with digestion.<\/span><\/li>\n<\/ol>\n<p>&nbsp;<\/p>\n<p><span style=\"color: #307897\"><a style=\"color: #307897\" href=\"https:\/\/lab.research.sickkids.ca\/sparc-molecular-analysis\/wp-content\/uploads\/sites\/61\/2017\/12\/Acetone-Precipitation.pdf\"><em><strong>Download and save a PDF copy of the acetone precipitation protocol. <\/strong><\/em><\/a><\/span><\/p>\n","protected":false},"excerpt":{"rendered":"<p>PROTOCOL ACETONE PRECIPITATION OF PROTEINS This protocol can be used to precipitate proteins from solutions where heavy detergents were used to prepare protein extracts. Required materials: Cold (-20\u00b0C) acetone (volume = 4X that of the protein samples to be precipitated). Centrifuge tubes made of acetone-compatible polypropylene and able to hold 5X sample volume. Centrifuge and&hellip;<\/p>\n","protected":false},"author":205,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-1790","page","type-page","status-publish","hentry","description-off"],"yoast_head":"<!-- This site is optimized with the Yoast SEO Premium plugin v27.0 (Yoast SEO v27.0) - https:\/\/yoast.com\/product\/yoast-seo-premium-wordpress\/ -->\n<title>Acetone Precipitation - SPARC BioCentre Molecular Analysis<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/lab.research.sickkids.ca\/sparc-molecular-analysis\/acetone-precipitation\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Acetone Precipitation\" \/>\n<meta property=\"og:description\" content=\"PROTOCOL ACETONE PRECIPITATION OF PROTEINS This protocol can be used to precipitate proteins from solutions where heavy detergents were used to prepare protein extracts. 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Required materials: Cold (-20\u00b0C) acetone (volume = 4X that of the protein samples to be precipitated). Centrifuge tubes made of acetone-compatible polypropylene and able to hold 5X sample volume. 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