{"id":266,"date":"2015-01-12T14:34:47","date_gmt":"2015-01-12T19:34:47","guid":{"rendered":"http:\/\/www.lifetein.com\/blog\/?p=266"},"modified":"2024-11-16T22:39:18","modified_gmt":"2024-11-17T03:39:18","slug":"lifetein-peptides-used-for-pulldown-assay","status":"publish","type":"post","link":"https:\/\/www.lifetein.com\/blog\/lifetein-peptides-used-for-pulldown-assay\/","title":{"rendered":"LifeTein Peptides Used for Pulldown Assay"},"content":{"rendered":"LifeTein synthesized multiple peptides for the pulldown experiments. The peptides were synthesized with biotin on the N terminus and\u00a0an aminohexanoic acid linker. Two types of peptides were designed: peptide without acetylation and peptide with multiple lysines (up to five sites) were acetylated in the synthetic peptides.\n

Pulldown Experiments With LiteTein Peptides<\/h2>\nThese peptides were bound to streptavidin-agarose beads and used for pulldown experiments using cell lysates. \u00a0Peptides, streptavidin agarose beads, and cell lysates were permitted to bind for overnight at 4 \u00b0C, \u00a0then beads were pelleted, washed five\u00a0times using spin columns, and proteins were eluted\u00a0in sample buffer and analyzed by SDS-PAGE and\u00a0immunoblotting.\n\nAll synthetic peptides had an N-terminal biotin. After incubation, peptides were spotted on nitrocellulose membranes and immunoblotted to detect acetylated Lysine (AcK) or total biotinylated peptide.\u00a0Data is shown below.\n\n
\"Acetylated<\/a>

Dot blot for biotin peptide with pulldown assay<\/p><\/div>\n\nReference:\n\nAcetylation of TUG Protein Promotes the Accumulation of GLUT4 Glucose Transporters in an Insulin-Responsive Intracellular Compartment, The Journal of Biological Chemistry, January 5, 2015, doi: 10.1074\/jbc.M114.603977 jbc.M114.603977.<\/a>\n

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LifeTein synthesized multiple peptides for the pulldown experiments. The peptides were synthesized with biotin on the N terminus and\u00a0an aminohexanoic acid linker. Two types of peptides were designed: peptide without acetylation and peptide with multiple lysines (up to five sites) … Continue reading →<\/span><\/a><\/p>\n","protected":false},"author":1,"featured_media":546,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"_crdt_document":"","_monsterinsights_skip_tracking":false,"_monsterinsights_sitenote_active":false,"_monsterinsights_sitenote_note":"","_monsterinsights_sitenote_category":0,"footnotes":""},"categories":[4],"tags":[50,45,46,49,48,47],"class_list":["post-266","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-peptide_synthesis","tag-biotin-peptide","tag-dot-blot","tag-immunoprecipitation","tag-peptide-acetylation","tag-peptide-pulldown","tag-western-blot"],"aioseo_notices":[],"_links":{"self":[{"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/posts\/266","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/comments?post=266"}],"version-history":[{"count":0,"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/posts\/266\/revisions"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/media\/546"}],"wp:attachment":[{"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/media?parent=266"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/categories?post=266"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.lifetein.com\/blog\/wp-json\/wp\/v2\/tags?post=266"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}