{"id":20144,"date":"2019-10-16T14:31:07","date_gmt":"2019-10-16T18:31:07","guid":{"rendered":"https:\/\/biochimie.umontreal.ca\/?page_id=20144"},"modified":"2019-11-04T13:34:41","modified_gmt":"2019-11-04T18:34:41","slug":"sec-mals","status":"publish","type":"page","link":"https:\/\/biochimie.umontreal.ca\/en\/scientific-platforms-bmm\/biophysics\/sec-mals\/","title":{"rendered":"SEC-MALS"},"content":{"rendered":"<p><div class=\"one-third first \">\n<a href=\"https:\/\/biochimie.umontreal.ca\/wp-content\/uploads\/sites\/37\/2018\/11\/sec-mals2.png\"><img loading=\"lazy\" decoding=\"async\" width=\"220\" height=\"159\" class=\"alignnone wp-image-18574\" alt=\"\" src=\"https:\/\/biochimie.umontreal.ca\/wp-content\/uploads\/sites\/37\/2018\/11\/sec-mals2-450x325.png\" srcset=\"https:\/\/biochimie.umontreal.ca\/wp-content\/uploads\/sites\/37\/2018\/11\/sec-mals2-450x325.png 450w, https:\/\/biochimie.umontreal.ca\/wp-content\/uploads\/sites\/37\/2018\/11\/sec-mals2-610x440.png 610w, https:\/\/biochimie.umontreal.ca\/wp-content\/uploads\/sites\/37\/2018\/11\/sec-mals2.png 669w\" sizes=\"auto, (max-width: 220px) 100vw, 220px\" \/><\/a><\/p>\n<p><div class=\"mybutton goal   \"><a href=\"https:\/\/faces.ccrc.uga.edu\/login0.php?account=BISTROP\" target=\"_blank\" rel=\"noopener noreferrer\">Online reservation<\/a><\/div><\/div><br \/>\n<div class=\"two-third  \">\n<strong><\/strong><\/p>\n<p><strong>Contacts<\/strong><\/p>\n<p><strong>Normand Cyr<\/strong><br \/>\nStructural biology platform manager<br \/>\n<a href=\"mailto:normand.cyr@umontreal.ca\">normand.cyr@umontreal.ca<\/a><br \/>\n514-343-6322<\/p>\n<p><strong>Pascale Legault<\/strong><br \/>\nProfessor<br \/>\n<a href=\"mailto:pascale.legault@umontreal.ca\">pascale.legault@umontreal.ca<\/a><br \/>\n<\/div><\/p>\n<div class=\"togglecontainer toggle_close_all \">\n\n<div class=\"toggler extralight-border\"><span class=\"text-title\"><strong>Description of the technique<\/strong><\/span><span class=\"toggle_icon extralight-border\"><span class=\"vert_icon extralight-border\"><\/span><span class=\"hor_icon extralight-border\"><\/span><\/span><\/div>\n<div class=\"toggle_wrap\">\n<div class=\"toggle_content\">\n<p style=\"text-align: justify\">Size Exclusion Chromatography with Multi-Angle Light Scattering analysis (SEC-MALS) is an absolute technique for the determination of the molar mass and the average radius of gyration (R<sub>g<\/sub>) of biomolecules in solution\u00a0following a separation by liquid chromatography. The technique uses static diffusion of a high intensity light source through a solution containing macromolecules. The intensity of this diffusion is then detected under several\u00a0angles, hence being able to calculate precisely the molar mass and the average radius of gyration (R<sub>g<\/sub>).<\/p>\n\n<\/div>\n<\/div>\n\n<div class=\"toggler extralight-border\"><span class=\"text-title\"><strong>Description of the equipment<\/strong><\/span><span class=\"toggle_icon extralight-border\"><span class=\"vert_icon extralight-border\"><\/span><span class=\"hor_icon extralight-border\"><\/span><\/span><\/div>\n<div class=\"toggle_wrap\">\n<div class=\"toggle_content\">\n<h3>Analytical chromatography (SEC)<\/h3>\n<p style=\"text-align: justify\">An A\u0308KTAmicro (GE Healthcare) liquid chromatography system is used to perform a high resolution separation of macromolecules (proteins, RNA, DNA) at analytical levels. The precision of low flow rate pumps enables the use of analytical-grade chromatography columns and injection of small quantities of material to be analyzed.<\/p>\n<h3 style=\"text-align: justify\">Multi-Angle Light Scattering (MALS) Detector<\/h3>\n<p style=\"text-align: justify\">A Dawn HELEOS II (Wyatt Technology) multi-angle light scattering detector is connected directly at the outlet of the chromatography column. The presence of multiple light scattering detectors enables the measurement of the light intensity at 18 different angles, offering great sensitivity (as low as 200 ng of BSA in solution) and both a wide range of molecular weights (200 Da to 1000 MDa) and of size (R<sub>g<\/sub> 10 to 500 nm).<\/p>\n<h3 style=\"text-align: justify\">Refractive Index Detector (RI)<\/h3>\n<p style=\"text-align: justify\">Finally, a OptiLab T-rEX (Wyatt Technology) refractive index detector is positionned downstream of the MALS detector in order to determine the concentration of biomolecules being separated.<\/p>\n<h3>Available chromatography columns<\/h3>\n<p>Several analytical liquid chromatography columns are available for SEC-MALS analysis.<\/p>\n<h4>Size-exclusion columns (SEC)<\/h4>\n<ul>\n<li>Superdex 200 Increase, 10&#215;300<\/li>\n<li>Superose 12, 3.2&#215;300<\/li>\n<li>Superdex 200 Increase, 3.2&#215;300<\/li>\n<li>Superdex 75, 3.2&#215;300<\/li>\n<li>Superose 6 Increase, 3.2&#215;300<\/li>\n<\/ul>\n<h4>Ion-exchange columns (IEX)<\/h4>\n<ul>\n<li>Mono S, 1.6&#215;5<\/li>\n<li>Mono Q, 1.6&#215;5<\/li>\n<\/ul>\n<h2>Softwares<\/h2>\n<ul>\n<li>ASTRA 6.1.6.5 (Wyatt Technology)<\/li>\n<\/ul>\n\n<\/div>\n<\/div>\n<br \/>\n\n<\/div>\n\n<p>&nbsp;<\/p>\n","protected":false},"excerpt":{"rendered":"<p>&nbsp;<\/p>\n","protected":false},"author":93,"featured_media":0,"parent":20142,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-20144","page","type-page","status-publish","hentry"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v25.0 - 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