{"id":3717,"date":"2019-05-29T05:25:38","date_gmt":"2019-05-29T09:25:38","guid":{"rendered":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/?p=3717"},"modified":"2021-06-17T11:34:14","modified_gmt":"2021-06-17T15:34:14","slug":"fluorophoreselection","status":"publish","type":"post","link":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/technical-tips\/fluorophoreselection\/","title":{"rendered":"Fluorophore selection and panel building &#8211; Spectra Viewer"},"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\/3717?pdf=3717\" 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<style>#blogpage-content h3{font-size:17.5px;color:#009fe3}#blogpage-content h4{color:#009453;margin-top:20px}#blogpage-content .btn-sq{color:#fff;font-size:.9rem;font-weight:600}#blogpage-content .btn-sq:hover{text-decoration:none;color:#ed7004}.btn-container{display:-webkit-box;display:-ms-flexbox;display:flex;-webkit-box-pack:justify;-ms-flex-pack:justify;justify-content:space-between;margin:1.3rem 0}.btn-sq .fad,.btn-sq .fal,.btn-sq .far,.btn-sq .fas{margin-right:10px}.btn-sq{border:2px solid #ed7004;padding:7px 13px;background:#ed7004;cursor:pointer;text-align:center;line-height:1.5rem;margin:0 auto;display:block}.btn-sq:hover{background:#fff}.vertical-btns{display:block}.vertical-btns .btn-sq{margin:1rem auto}@media (max-width:768px){#blogpage-content .btn-sq{font-size:1.05rem}}#blogpage-content figure{margin:0 auto;margin-bottom:2rem;border:1px solid #ccc;width:70%}#blogpage-content figure img{display:block;margin:0 auto;width:100%}#blogpage-content figure figcaption{font-size:.875rem;line-height:1.35rem;padding:10px;color:#222}#blogpage-content figure a{font-size:.875rem}<\/style>\n<p><script src=\"https:\/\/kit.fontawesome.com\/904923013f.js\" crossorigin=\"anonymous\"><\/script><\/p>\n<div id=\"blogpage-content\">\n<h2>Fluorescent probes or fluorophores (fluorescent dyes or proteins) are coupled to a secondary antibody or streptavidin to allow visualization of an analyte. Each fluorophore has its specific spectral characteristics, with excitation and emission spectra particular to the molecule.&nbsp; Use the spectra viewer to build dye panels and compare the suitability of dyes for your application.<\/h2>\n<p><span style=\"font-weight: 400;\">Fluorescent conjugates facilitate the simultaneous detection of multiple analytes for a number of techniques including flow cytometry, fluorescence microscopy (epifluorescence, confocal, multiphoton and super-resolution techniques), Western blotting and ELISA. Each fluorophore has its own spectral characteristics, with excitation and emission spectra particular to the molecule. Use the spectra viewer to build dye panels suitable for your experiment by plotting individual dye characteristics and instrument set up.&nbsp; For multiple labeling protocols, the dye panel choices&nbsp;are constrained by instrumentation and sample specifics such as lasers and filters available. To achieve good color separation, choose fluorophores with minimal spectral overlap.<\/span><\/p>\n<figure><img decoding=\"async\" src=\"\/secondary-antibody-resource\/wp-content\/uploads\/spectral-viewer-1.jpg\" alt=\"spectral viewer\"><figcaption><strong>Fig 1:<\/strong> Use the Spectra Viewer to plot the spectral characteristics of dyes to build dye panels suitable for your application. The spectra viewer can be found on individual product pages and <a href=\"\/technical\/products\/conjugate-selection\/fluorophore-selection#spectra\">here<\/a>.<\/figcaption><\/figure>\n<div class=\"btn-container\"><a class=\"btn-sq\" href=\"\/technical\/products\/conjugate-selection\/fluorophore-selection#spectra\"><i class=\"fas fa-chart-area\"><\/i>Use the spectra Viewer<\/a><\/div>\n<h3>How to select Fluorophores<\/h3>\n<p>The choice of fluorescent probe depends on a number of experimental variables.<\/p>\n<h4>Instrument<\/h4>\n<p>Consider excitation capabilities (lamp, lasers or LEDs determine excitation wavelengths) and the number of&nbsp;channels and filters available. &nbsp;The user should review the parameters of the filter sets included with&nbsp;their instrument. Long-pass filters provide the best sensitivity, while narrow band-pass filters enable&nbsp;discrimination between signals from different fluorophores in multiple labeling applications.<br \/>\nUse our Spectra viewer to develop your panel and build dye panels specific to&nbsp;your instrument.<\/p>\n<h4>Degree of color separation required<\/h4>\n<p>To achieve good color separation in a multiple labeling protocol, choose fluorophores with minimal spectral overlap.<\/p>\n<h4>Experimental sample<\/h4>\n<p>Choose fluorophores whose spectra do not overlap with endogenous fluorescence. Examine an unstained specimen&nbsp;under available filter sets to determine whether the sample exhibits autofluorescence.<\/p>\n<p><strong>Find out more about fluorescent conjugates available from Jackson ImmunoResearch <a href=\"\/technical\/products\/conjugate-selection\">here<\/a>.<\/strong><\/p>\n<\/div>\n<p>&nbsp;<\/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=\"\/technical\/products\/protocols\/multiple-labeling\">Multiple labeling using Secondary Antibodies<\/a><\/td>\n<td class=\"span6\"><a href=\"\/catalog\/1\">Whole&nbsp;IgG affinity-purified antibodies<\/a><\/td>\n<\/tr>\n<tr>\n<td class=\"span6\"><a href=\"\/secondary-antibody-resource\/immuno-techniques\/direct-and-indirect-western-blotting\/\">Direct and Indirect Western blotting<\/a><\/td>\n<td class=\"span6\"><a href=\"\/technical\/products\/conjugate-selection#spectra\">Build dye panels &#8211; Spectra Viewer<\/a><\/td>\n<\/tr>\n<tr>\n<td class=\"span6\"><a href=\"\/technical\/products\/antibody-selection\">Choosing your Secondary Antibody<\/a><\/td>\n<td class=\"span6\"><a href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/VHH-Discovery\">Secondary Antibodies for VHH discovery<\/a><\/td>\n<\/tr>\n<\/tbody>\n<\/table>\n<p style=\"text-align: left;\">\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 Fluorescent probes or fluorophores (fluorescent dyes or proteins) are coupled to a secondary antibody or streptavidin to allow visualization of an analyte. Each fluorophore has its specific spectral characteristics, with excitation and emission spectra particular to the molecule.&nbsp; Use the spectra viewer to build dye panels and compare the suitability of dyes for [&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":3,"featured_media":3726,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"content-type":""},"categories":[16],"tags":[],"acf":[],"_links":{"self":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/3717"}],"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\/3"}],"replies":[{"embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/comments?post=3717"}],"version-history":[{"count":23,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/3717\/revisions"}],"predecessor-version":[{"id":8224,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/3717\/revisions\/8224"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/media\/3726"}],"wp:attachment":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/media?parent=3717"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/categories?post=3717"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/tags?post=3717"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}