{"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

Peptide Synthesis Home Page<\/a><\/p>\n\n\nOur Services:\n\n \n\nCOVID-19 Services & Products<\/a>\n\n \n\nCustom Antibody Services<\/a>\n\n \n\nRush Peptide Synthesis<\/a>\n\n \n\nPeptide Nucleic Acids (PNAs)<\/a>\n\n \n\nCustom Peptide Synthesis Services<\/a>\n\n \n\nGene Synthesis Service<\/a>\n\n \n\nCustom Chemical Synthesis<\/a>\n\n \n\nOther Posts:\n\n \n\nAmino acid composition of cell-penetrating peptides (CPPs)<\/a>\n\nTo Make Simpler and Better Biosensors<\/a>\n\nA Simple Protocol to Refold Peptides or Small Proteins<\/a>\n\nPeptide Applications<\/a>\n\n","protected":false},"excerpt":{"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) … 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":{"_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":[],"aioseo_head":"\n\t\t\n\t\n\t\n\t\n\t\n\t\n\t\n\t\n\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\t\t\n\n","aioseo_head_json":{"title":"LifeTein Peptides Used for Pulldown Assay | LifeTein Peptide Blog","description":"Biotin peptides were used for pulldown assay using streptavidin agaroses. The peptides were acetylated on multiple lysines to study the protein regulations.","canonical_url":"https:\/\/www.lifetein.com\/blog\/lifetein-peptides-used-for-pulldown-assay\/","robots":"max-image-preview:large","keywords":"peptide pulldown,peptide biotin,peptide acetylation,dot blot,immunoprecipitation,biotin peptide,western blot","webmasterTools":{"google-site-verification":"xKDW6DUiGuZ6nwglNeWlVWTgzTtMPzas13ROL63ytTc","msvalidate.01":"5B15601215D876311F84CFEFB0F72F18","miscellaneous":""},"schema":null,"og:locale":"en_US","og:site_name":"LifeTein Peptide Synthesis Blog","og:type":"activity","og:title":"LifeTein Peptides Used for Pulldown Assay | LifeTein Peptide Blog","og:description":"Biotin peptides were used for pulldown assay using streptavidin agaroses. The peptides were acetylated on multiple lysines to study the protein regulations.","og:url":"https:\/\/www.lifetein.com\/blog\/lifetein-peptides-used-for-pulldown-assay\/","fb:admins":"233983946638783","og:image":"https:\/\/www.lifetein.com\/blog\/wp-content\/uploads\/2020\/02\/LifeTein-Blog1-2-scaled.jpg","og:image:secure_url":"https:\/\/www.lifetein.com\/blog\/wp-content\/uploads\/2020\/02\/LifeTein-Blog1-2-scaled.jpg","twitter:card":"summary","twitter:site":"@lifetein","twitter:title":"LifeTein Peptides Used for Pulldown Assay | LifeTein Peptide Blog","twitter:description":"Biotin peptides were used for pulldown assay using streptavidin agaroses. The peptides were acetylated on multiple lysines to study the protein regulations.","twitter:creator":"@lifetein","twitter:image":"https:\/\/www.lifetein.com\/blog\/wp-content\/uploads\/2020\/02\/LifeTein-Blog1-2-scaled.jpg"},"aioseo_meta_data":{"post_id":"266","title":"LifeTein Peptides Used for Pulldown Assay | #site_title","description":"Biotin peptides were used for pulldown assay using streptavidin agaroses. The peptides were acetylated on multiple lysines to study the protein regulations.","keywords":[{"label":"peptide pulldown","value":"peptide pulldown"},{"label":"peptide biotin","value":"peptide biotin"},{"label":"peptide acetylation","value":"peptide acetylation"},{"label":"dot blot","value":"dot blot"},{"label":"immunoprecipitation","value":"immunoprecipitation"}],"keyphrases":{"focus":{"keyphrase":"","score":0,"analysis":{"keyphraseInTitle":{"score":0,"maxScore":9,"error":1}}},"additional":[]},"primary_term":null,"canonical_url":null,"og_title":null,"og_description":null,"og_object_type":"activity","og_image_type":"default","og_image_url":null,"og_image_width":null,"og_image_height":null,"og_image_custom_url":null,"og_image_custom_fields":null,"og_video":"","og_custom_url":null,"og_article_section":null,"og_article_tags":null,"twitter_use_og":true,"twitter_card":"summary","twitter_image_type":"default","twitter_image_url":null,"twitter_image_custom_url":null,"twitter_image_custom_fields":null,"twitter_title":null,"twitter_description":null,"schema":{"blockGraphs":[],"customGraphs":[],"default":{"data":{"Article":{"id":"aioseo-article-633cec9287827","slug":"article","graphName":"Article","label":"Article","properties":{"type":"BlogPosting","name":"#post_title","headline":"#post_title","description":"#post_excerpt","image":"","keywords":"","author":{"name":"#author_name","url":"#author_url"},"dates":{"include":true,"datePublished":"","dateModified":""}}},"Course":[],"Dataset":[],"FAQPage":[],"Movie":[],"Person":[],"Product":[],"ProductReview":[],"Car":[],"Recipe":[],"Service":[],"SoftwareApplication":[],"WebPage":[]},"graphName":"Article","isEnabled":true},"graphs":[]},"schema_type":"default","schema_type_options":"{\"article\":{\"articleType\":\"BlogPosting\"},\"course\":{\"name\":\"\",\"description\":\"\",\"provider\":\"\"},\"faq\":{\"pages\":[]},\"product\":{\"reviews\":[]},\"recipe\":{\"ingredients\":[],\"instructions\":[],\"keywords\":[]},\"software\":{\"reviews\":[],\"operatingSystems\":[]},\"webPage\":{\"webPageType\":\"WebPage\"}}","pillar_content":false,"robots_default":true,"robots_noindex":false,"robots_noarchive":false,"robots_nosnippet":false,"robots_nofollow":false,"robots_noimageindex":false,"robots_noodp":false,"robots_notranslate":false,"robots_max_snippet":"-1","robots_max_videopreview":"-1","robots_max_imagepreview":"none","priority":null,"frequency":"default","location":null,"local_seo":null,"seo_analyzer_scan_date":null,"breadcrumb_settings":null,"limit_modified_date":false,"open_ai":"{\"title\":{\"suggestions\":[],\"usage\":0},\"description\":{\"suggestions\":[],\"usage\":0}}","ai":null,"created":"2020-12-04 22:27:49","updated":"2025-06-18 02:41:15"},"aioseo_breadcrumb":"

\n\t\t\tHome<\/a>\n\t\t<\/span>»<\/span>\n\t\t\tPeptide<\/a>\n\t\t<\/span>»<\/span>\n\t\t\tLifeTein Peptides Used for Pulldown Assay\n\t\t<\/span><\/div>","aioseo_breadcrumb_json":[{"label":"Home","link":"https:\/\/www.lifetein.com\/blog"},{"label":"Peptide","link":"https:\/\/www.lifetein.com\/blog\/category\/peptide_synthesis\/"},{"label":"LifeTein Peptides Used for Pulldown Assay","link":"https:\/\/www.lifetein.com\/blog\/lifetein-peptides-used-for-pulldown-assay\/"}],"_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}]}}