{"id":8247,"date":"2021-07-20T04:53:28","date_gmt":"2021-07-20T08:53:28","guid":{"rendered":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/?p=8247"},"modified":"2021-07-20T05:01:24","modified_gmt":"2021-07-20T09:01:24","slug":"how_can-his-tags_benefit_your_research","status":"publish","type":"post","link":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/company-news\/how_can-his-tags_benefit_your_research\/","title":{"rendered":"How can His-Tags Benefit your Research?"},"content":{"rendered":"\n\n\n\t<div class=\"dkpdf-button-container\" style=\" text-align:right \">\n\n\t\t<a class=\"dkpdf-button\" href=\"\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/8247?pdf=8247\" target=\"_blank\"><span class=\"dkpdf-button-icon\"><i class=\"fa fa-file-pdf-o\"><\/i><\/span> Download PDF<\/a>\n\n\t<\/div>\n\n\n\n\n\n<p><style>.entry h3{font-size:17.5px;color:#009fe3}.entry h4{font-size:16px;color:#009453}.btn-sq{color:#fff;font-size:.9rem;font-weight:600;border:2px solid rgb(237, 112, 4);padding:7px 13px;background:rgb(237, 112, 4);cursor:pointer;text-align:center;line-height:1.5rem;margin:0 auto;}.btn-sq:hover{text-decoration:none;color:rgb(237, 112, 4);background:#fff;}.btn-container{display:flex;width:100%;margin:1.3rem 0;}.btn-container .fad{margin-right:10px;}@media(max-width: 768px){.btn-sq{font-size:1.05rem;}}<\/style><\/p>\n<p><script src=\"https:\/\/kit.fontawesome.com\/904923013f.js\" crossorigin=\"anonymous\"><\/script><\/p>\n<h2><span style=\"font-weight: 400;\">Protein tags underpin a broad range of research applications. These include affinity purification, where protein tags enable recombinant fusion proteins to be captured on a solid matrix, and immunodetection techniques such as Western blot, ELISA, and immunohistochemistry (IHC), where anti-tag antibodies are used for target detection. Of the many different protein tags that are incorporated into recombinant protein constructs, His-tags are popular for their small size. Not only does this prevent them from interfering with protein structure or function, but it also provides several further advantages.<\/span><\/h2>\n<p><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter wp-image-4121\" style=\"width: 100%;\" src=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/block-flow-image.jpg\" alt=\"\" width=\"600\" height=\"440\" srcset=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/block-flow-image.jpg 412w, https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/block-flow-image-300x220.jpg 300w\" sizes=\"(max-width: 600px) 100vw, 600px\" \/><\/p>\n<h3>What are protein tags?<\/h3>\n<p><span style=\"font-weight: 400;\">Broadly speaking, the term protein tag can be used to describe any protein that is incorporated into another protein to enable its isolation or detection. Commonly used tags include glutathione S-transferase (GST), maltose-binding protein (MBP), thioredoxin, green fluorescent protein (GFP), chitin-binding domain (CBD), to name but a few. Additionally, tags comprising just a short sequence of amino acids are also popular, with well-known examples being His, Myc, FLAG\u00ae, HA, and Strep-tag. Protein tags are typically expressed either at the N-terminus or C-terminus of a recombinant protein. However, for certain research applications, a different tag may be expressed at each terminus; for example, to facilitate the detection of a cleavage product.<\/span><\/p>\n<h3>What are the advantages of His-tags?<\/h3>\n<p><span style=\"font-weight: 400;\">His-tags, also known as poly-histidine tags, comprises of a series of six to ten histidine residues, with the 6-residue tag (HHHHHH, referred to as 6-His) being used most often. A main advantage of His-tags is their small size; 6-His has a molecular weight of just 1 kDa. This makes them more robust than larger protein tags as well as prevents them from interfering with protein folding. Notably, His-tags enable affinity purification using inexpensive and widely available nickel or cobalt resins, unlike proprietary tags such as FLAG that require dedicated resins for protein purification. Moreover, His-tags have relatively low immunogenicity, eliminating the need for tag removal before using purified recombinant proteins as immunogens during antibody production.<\/span><\/p>\n<h3>How are His-tags used?<\/h3>\n<p><span style=\"font-weight: 400;\">His-tags are often used to identify expressing cells when producing recombinant proteins. For example, using anti-His antibodies, researchers can easily detect a secreted His-tagged protein in cell culture supernatant when selecting suitable clones for expansion. His-tags are also widely exploited for affinity purification, where they enable target proteins to be captured from solution; after protein binding, via the His-tag, to a resin, any unbound components are washed away and the His-tagged protein is then eluted.<\/span><\/p>\n<p><span style=\"font-weight: 400;\">Another use of His-tags is to facilitate immunodetection via techniques such as Western blot, ELISA, and IHC. This involves either using labeled anti-His primary antibodies for direct detection of protein targets, or combining unlabeled anti-His antibodies with labeled secondary antibodies during indirect detection; critically, the latter approach provides signal amplification that can enhance the detection of rare targets.<\/span><\/p>\n<p><span style=\"font-weight: 400;\">Importantly, detecting His-tags rather than the target protein itself reduces the need to use expensive, target-specific antibodies for immunodetection. This can especially be beneficial when working with a novel target for which few antibodies are available, or when designing a multiplexed experiment that requires maximum flexibility in the choice of host species. Furthermore, using a His-tag for identification purposes ensures target protein epitopes remain free to bind target-specific antibodies.<\/span><\/p>\n<p><span style=\"font-weight: 400;\">Jackson ImmunoResearch specializes in producing secondary antibodies for life science applications. Our portfolio includes a broad range of secondaries conjugated to<\/span><a href=\"https:\/\/www.jacksonimmuno.com\/catalog\/1\"> <span style=\"font-weight: 400;\">biotin<\/span><\/a><span style=\"font-weight: 400;\">,<\/span><a href=\"https:\/\/www.jacksonimmuno.com\/catalog\/1\"> <span style=\"font-weight: 400;\">HRP<\/span><\/a><span style=\"font-weight: 400;\">, and fluorescent dyes and proteins such as Alexa Fluors\u00ae,<\/span><a href=\"https:\/\/www.jacksonimmuno.com\/catalog\/4\"> <span style=\"font-weight: 400;\">R-PE, APC, and PerCP<\/span><\/a><span style=\"font-weight: 400;\">. We have also recently expanded our product offering with the launch of our<\/span><a href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/groups\/anti-his\"> <span style=\"font-weight: 400;\">NEW Anti-His Tag Polyclonal Antibody<\/span><\/a><span style=\"font-weight: 400;\">. Every one of our antibody reagents is provided with a suggested working concentration range for each application in which it has been validated and is backed by expert technical support for results you can trust.<\/span><\/p>\n<p><a href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/groups\/anti-his\"><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter size-full wp-image-8274\" src=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Anti-his-A.jpg\" alt=\"\" width=\"729\" height=\"90\" srcset=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Anti-his-A.jpg 729w, https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Anti-his-A-300x37.jpg 300w\" sizes=\"(max-width: 729px) 100vw, 729px\" \/><\/a><\/p>\n<table class=\"table blogLinks aligncenter\">\n<thead>\n<tr>\n<th class=\"span6\">Learn more:<\/th>\n<th class=\"span6\">Do more:<\/th>\n<\/tr>\n<\/thead>\n<tbody>\n<tr>\n<td class=\"span6\"><a href=\"\/secondary-antibody-resource\/immuno-techniques\/colorimetric-western-blotting\/\">Colorimetric western blotting<\/a><\/td>\n<td class=\"span6\"><strong><a href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/conjugate-selection\/fluorophore-selection#spectra\">Spectra Viewer<\/a><\/strong><\/td>\n<\/tr>\n<tr>\n<td class=\"span6\"><a href=\"\/secondary-antibody-resource\/immuno-techniques\/fluorescent-western-blotting\/\">Chemiluminescence western blotting<\/a><\/td>\n<td class=\"span6\"><a href=\"\/technical\/products\/groups\/signal-enhancement\">Antibodies for signal enhancement<\/a><\/td>\n<\/tr>\n<tr>\n<td class=\"span6\" colspan=\"2\"><a href=\"\/secondary-antibody-resource\/immuno-techniques\/fluorescent-western-blotting\/\">Fluorescent western blotting<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n\n\n<p><\/p>\n<!-- AddThis Advanced Settings generic via filter on the_content --><!-- AddThis Share Buttons generic via filter on the_content --><!-- AddThis Related Posts generic via filter on the_content -->","protected":false},"excerpt":{"rendered":"<p>Download PDF Protein tags underpin a broad range of research applications. These include affinity purification, where protein tags enable recombinant fusion proteins to be captured on a solid matrix, and immunodetection techniques such as Western blot, ELISA, and immunohistochemistry (IHC), where anti-tag antibodies are used for target detection. Of the many different protein tags that [&hellip;]<!-- AddThis Advanced Settings generic via filter on get_the_excerpt --><!-- AddThis Share Buttons generic via filter on get_the_excerpt --><!-- AddThis Related Posts generic via filter on get_the_excerpt --><\/p>\n","protected":false},"author":11,"featured_media":4140,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"content-type":""},"categories":[19,20],"tags":[],"acf":[],"_links":{"self":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/8247"}],"collection":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/users\/11"}],"replies":[{"embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/comments?post=8247"}],"version-history":[{"count":6,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/8247\/revisions"}],"predecessor-version":[{"id":8278,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/8247\/revisions\/8278"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/media\/4140"}],"wp:attachment":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/media?parent=8247"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/categories?post=8247"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/tags?post=8247"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}