{"id":351,"date":"2021-04-12T10:50:09","date_gmt":"2021-04-12T10:50:09","guid":{"rendered":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/?page_id=351"},"modified":"2021-04-12T10:50:23","modified_gmt":"2021-04-12T10:50:23","slug":"sample-optimisation","status":"publish","type":"page","link":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/sample-optimisation\/","title":{"rendered":"Sample Optimisation"},"content":{"rendered":"\n<div class=\"vf-grid | vf-grid__col-3\"><div class=\"vf-grid__col--span-2\"><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<p>Sample optimization is often a key step towards structure determination. Fine tuning of the sample preparation conditions can reduce heterogeneity and a &#8216;secret ingredient&#8217; may increase the stability of a protein. It is a good idea to combine preliminary results from structural studies with the results obtained from the quality control measurements as a basis to optimise the sample conditions. The SPC facility provides an integrated package of quality control coupled to sample optimization.<\/p>\n\n\n\n<h2 class=\"wp-block-heading\"><strong>Thermofluor (DSF) optimisation screen<\/strong><\/h2>\n\n\n\n<p>A screen of 96 different conditions varying buffers, salts and a range of additives are tested to see whether the protein becomes more stable as reflected by the melting curve. Details and prices on request (<a href=\"mailto:spc@embl-hamburg.de\">spc@embl-hamburg.de<\/a>).<\/p>\n\n\n\n<h2 class=\"wp-block-heading\"><strong>nano Differential scanning fluorimetry (nanoDSF)\u00a0optimisation screen<\/strong><\/h2>\n\n\n\n<p>In contrast to conventional DSF&nbsp;methods, nanoDSF uses tryptophan or tyrosin fluorescence to monitor protein unfolding. Both the fluorescence intensity and the fluorescence maximum strongly depends on the close surroundings of the tryptophan.<sup><\/sup>&nbsp;Therefore, the ratio of the fluorescence intensities at 350&nbsp;nm and 330&nbsp;nm is suitable to detect any changes in protein structure, for example due to protein unfolding.<\/p>\n\n\n\n<p>Using the 96 different conditions varying buffers, salts and a range of additives on the nanoDSF,&nbsp;optimal buffer components can be identified for each individual protein for further purification,&nbsp;protein buffer optimization, identifying optimal conditions for storage, assay screening, and crystallization.&nbsp;Details and prices on request (<a href=\"mailto:spc@embl-hamburg.de\">spc@embl-hamburg.de<\/a>).<\/p>\n\n<\/div>\n<\/div>\n\n\n<div><!--[vf\/content]-->\n<div class=\"vf-content\">\n\n<div class=\"wp-block-image\"><figure class=\"vf-figure  | vf-figure--align vf-figure--align-inline-start   size-full is-resized\"><img loading=\"lazy\" decoding=\"async\" class=\"vf-figure__image\" src=\"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-content\/uploads\/2021\/04\/biomekoptimisation.jpg\" alt=\"\" class=\"wp-image-352\" width=\"375\" height=\"250\" srcset=\"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-content\/uploads\/2021\/04\/biomekoptimisation.jpg 500w, https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-content\/uploads\/2021\/04\/biomekoptimisation-300x200.jpg 300w\" sizes=\"auto, (max-width: 375px) 100vw, 375px\" \/><\/figure><\/div>\n\n<\/div>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"template-title-left-aligned.php","meta":{"_acf_changed":false,"footnotes":""},"embl_taxonomy":[],"class_list":["post-351","page","type-page","status-publish","hentry"],"acf":[],"embl_taxonomy_terms":[],"_links":{"self":[{"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/pages\/351","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/comments?post=351"}],"version-history":[{"count":2,"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/pages\/351\/revisions"}],"predecessor-version":[{"id":354,"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/pages\/351\/revisions\/354"}],"wp:attachment":[{"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/media?parent=351"}],"wp:term":[{"taxonomy":"embl_taxonomy","embeddable":true,"href":"https:\/\/www.embl.org\/groups\/sample-preparation-characterisation\/wp-json\/wp\/v2\/embl_taxonomy?post=351"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}