{"id":10546,"date":"2023-11-30T10:17:14","date_gmt":"2023-11-30T15:17:14","guid":{"rendered":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/?p=10546"},"modified":"2023-11-30T10:17:18","modified_gmt":"2023-11-30T15:17:18","slug":"popular-bead-based-multiplex-assay","status":"publish","type":"post","link":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/immuno-techniques\/popular-bead-based-multiplex-assay\/","title":{"rendered":"Bead-based multiplexing assays: Luminex\u00ae xMAP\u00ae Technology"},"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\/10546?pdf=10546\" 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>.entry p, .entry ol{font-size: 1rem;}.entry h3{color:#009fe3;margin-top:0}.entry h4{color:#009453;margin-top:1.5rem}.entry h5{color: #003a5c; font-size: 1.05rem;margin-top:1.5rem;}.entry figure{margin:32px auto;border:1px solid #ccc}.entry figure img{width:100%;display:block;margin:0 auto}.entry figcaption{font-size:.875rem;line-height:1.35rem;padding:10px;color:#222}.entry .blog-tbl{margin:1rem auto;caption-side:bottom}.entry .blog-tbl td,.entry .blog-tbl th{padding:10px 9px;border:1px solid #00172b;text-align:left}.entry .blog-tbl td{border-color:#003a5c}.entry .blog-tbl th{background-color:#00172b;color:#fff;border-left-color:#fff;border-right-color:#fff}.entry .blog-tbl th p{color:#fff}.entry .blog-tbl th:first-of-type{border-left-color:#00172b}.entry .blog-tbl th:last-of-type{border-right-color:#00172b}.entry .blog-tbl p{text-align:left;margin:0}.entry .box-note{border:2px solid #009453;padding:12px;margin:1rem 0}.entry .box-note p{margin:0;padding:0}.entry .styled-list{list-style-type:none}.entry .styled-list li{margin-top:1rem;line-height:1rem;font-size:1rem;}.entry .styled-list li::before{font-family:\"Font Awesome 6 Pro\";display:inline-block;content:\"\\f3c5\";-webkit-transform:rotate(-90deg);transform:rotate(-90deg);margin-left:-20px;margin-right:11px;font-size:.75rem;color:#ed7004;font-weight:600}.entry .styled-list ol li::before{display:none}.entry .styled-list li>ul li::before{font-weight:200}.entry .overview{width:-webkit-fit-content;width:-moz-fit-content;width:fit-content;padding:16px;margin:1rem auto;border:1px solid #eee}.entry .overview hr{margin-top:14px}.entry .overview-text{text-align:center;font-size:1rem;margin:0}.entry .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;}.entry .btn-sq:hover{text-decoration:none;color:rgb(237, 112, 4);background:#fff;}.entry .btn-container{display:flex;width:100%;margin:1.3rem 0;}.entry .btn-container .fa-solid, .entry .btn-container .fa-duotone{margin-right:10px;}@media(max-width: 768px){.entry .btn-sq{font-size:1.05rem;}}.entry .footnote ul {margin-left: 0;list-style-type: none;}.entry .footnote ul li {font-size: .8rem;line-height: 1.2rem;margin-top: 0;}.entry .footnote ul li::before {display: none;}<\/style>\n<style>.entry .styled-list li{line-height:24px}<\/style>\n<p><script src=\"https:\/\/kit.fontawesome.com\/904923013f.js\" crossorigin=\"anonymous\"><\/script><\/p>\n<style>.entry figure:first-of-type{max-width: 420px;}<\/style>\n<style>@media (max-width: 980px) {.entry .btn-container{display:block;}.entry .btn-container .btn-sq{width:300px}}<\/style>\n<h2>According to Luminex\u00ae (a DiaSorin company), xMAP\u00ae is the world<span style=\"font-weight: 400;\">\u2032<\/span>s most used multiplexing technology, \u00a0a claim that is backed by over 70,000 peer-reviewed publications. These include multiple references to serology assays developed using Jackson ImmunoResearch secondary antibodies, which serve as essential detection reagents for many diagnostic assays using this platform.<\/h2>\n<h4><strong>Principles of the Luminex\u00ae assay<span style=\"font-weight: 400;\">\u2032<\/span><\/strong><\/h4>\n<p>xMAP\u00ae Technology is based on the use of color-coded beads that are dyed with different ratios of two fluorophores (one red and one infrared) to provide a unique spectral signature. In a typical assay setup, each spectrally distinct bead set is coated with a different analyte-specific antibody and is initially blocked prior to incubation with the sample. After washing to remove any unbound material, biotinylated analyte-specific antibodies are added to form a sandwich complex before a streptavidin-phycoerythrin (PE) conjugate completes the assemblage. The beads are then read on a flow cytometry-based Luminex\u00ae instrument, where a red (630 nm) laser is used to identify each bead set and a green laser (532 nm) determines the magnitude of the PE-derived signal, which is directly proportional to the amount of bound analyte. This setup is shown in Figure 1.<\/p>\n<figure><img decoding=\"async\" loading=\"lazy\" class=\"wp-image-10558 size-full aligncenter\" src=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Streptavidin-Illustrations-23-0911_01-min.jpg\" alt=\"\" width=\"450\" height=\"352\" srcset=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Streptavidin-Illustrations-23-0911_01-min.jpg 450w, https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Streptavidin-Illustrations-23-0911_01-min-300x235.jpg 300w\" sizes=\"(max-width: 450px) 100vw, 450px\" \/><figcaption><strong>Figure 1:<\/strong> A typical Luminex\u00ae (xMAP\u00ae) assay setup.<br \/>\nAn alternative approach involves coating each bead set with a protein analyte to capture antibodies from sample types including serum, saliva, and cerebrospinal fluid. In this setup, detection is usually accomplished using a PE-conjugated anti-species antibody (Figure 2a) or a biotinylated anti-species antibody in combination with a streptavidin-PE conjugate. Secondary antibodies from Jackson ImmunoResearch are widely used for these types of assays.<\/figcaption><\/figure>\n<figure><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter wp-image-10568 size-full\" src=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Luminex-2a-2b-1.jpg\" alt=\"\" width=\"745\" height=\"383\" srcset=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Luminex-2a-2b-1.jpg 745w, https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Luminex-2a-2b-1-300x154.jpg 300w\" sizes=\"(max-width: 745px) 100vw, 745px\" \/><figcaption><strong>Figure 2:<\/strong> Alternative Luminex set-ups using analyte-bound beads to capture antibodies from patient samples and then detected using the following approaches. A. PE-conjugated anti-species antibody against sample species. B. biotinylated anti-species antibody followed by PE-conjugated Streptavidin to generate signal amplification.<\/figcaption><\/figure>\n<div class=\"btn-container\"><a class=\"btn-sq\" href=\"\/technical\/products\/groups\/streptavidin\"><i class=\"fa-solid fa-magnifying-glass\"><\/i>Browse Streptavidin<\/a><br \/>\n<a class=\"btn-sq\" href=\"\/technical\/products\/groups\/anti-human-secondary-antibodies\"><i class=\"fa-solid fa-magnifying-glass\"><\/i>Browse Anti-Human Antibodies<\/a><\/div>\n<hr \/>\n<h4><strong>Advantages of xMAP\u00ae Technology<\/strong><\/h4>\n<p>Like other multiplexed immunoassay formats, xMAP\u00ae Technology offers savings in terms of sample material, reagents, and time, as well as eliminates confounding variables like different buffer systems, ambient conditions, and operators for a more accurate comparison of different analytes within the same assay. However, a distinguishing feature of xMAP\u00ae Technology is that it can simultaneously detect up to 500 analytes per sample, which is more than most other multiplexed immunoassays. Additionally, it is now possible with the xMAP INTELLIFLEX\u00ae to measure two parameters per analyte. \u201cThe xMAP INTELLIFLEX\u00ae enhances the capabilities of the original xMAP technology by incorporating a second (405 nm) detection channel,\u201d explains Dominic Andrada, Scientific Applications Senior Marketing Manager at Luminex. \u201cThis enables researchers to effectively double the number of readouts obtained per sample. For example, using the xMAP INTELLIFLEX\u00ae, one can detect a protein of interest and a specific post-translational modification on the same bead.\u201d Another important advantage of xMAP\u00ae Technology is that the small size and low density of the beads equates to shorter assay times than ELISAs and microarrays, which are limited by solid-phase kinetics<sup>1<\/sup>.<\/p>\n<hr \/>\n<h4><strong>Applications of xMAP\u00ae Technology<\/strong><\/h4>\n<p>Because xMAP\u00ae Technology is highly flexible, it has utility for an almost infinite number of research applications. These include its use to study cancer (e.g., through measuring immune checkpoint proteins, circulating tumor markers, or markers of metastasis), inflammation (e.g., by quantifying cytokines, chemokines, or autoantibodies), and cell signaling pathways (e.g., via monitoring of Akt\/mTOR, NF\u03baB, or STAT signaling molecules). xMAP\u00ae Technology is also used for investigating the underlying mechanisms of metabolism, aging, and virology, as well as for studying neurodegeneration, metabolic disorders, and cardiovascular disease.<\/p>\n<hr \/>\n<h4><strong>High-quality reagents from Jackson ImmunoResearch for xMAP\u00ae assays<\/strong><\/h4>\n<p>Jackson ImmunoResearch specializes in producing secondary antibodies for life science applications and our products have been extensively cited for use with xMAP\u00ae Technology.<\/p>\n<p>Examples include<\/p>\n<ul>\n<li>2021 <em>Scientific Reports <\/em>publication in which our biotinylated donkey anti-human IgG Fc-specific antibody was used for detecting IgG antibodies to three noroviruses and <em>Cryptosporidium <\/em>in the saliva of individuals who reported swallowing water from Lake Michigan.<\/li>\n<li>2023 <em>Immunology <\/em>article citing our PE-labeled goat anti-human IgG antibody for detecting anti-SARS-CoV-2 antibodies in serum<sup>2,3<\/sup>.<\/li>\n<li>R-Phycoerythrin AffiniPure F(ab<span style=\"font-weight: 400;\">\u2032<\/span>)\u2082 Fragment Goat Anti-Mouse IgM, \u00b5 chain specific and R-Phycoerythrin AffiniPure F(ab<span style=\"font-weight: 400;\">\u2032<\/span>)\u2082 Fragment Goat Anti-Mouse IgG, Fc\u03b3 fragment specific antibodies featured in an <em>SLAS <\/em>publication reporting the development of an xMAP\u00ae assay for detecting anti-glycan antibodies in plasma, ascites, and cerebrospinal fluid<sup>4<\/sup>.<\/li>\n<\/ul>\n<p>Changes in glycosylation have been linked to congenital disorders, autoimmune diseases, and cancer, and it is thought that generating anti-glycan antibody profiles from different human body fluids may help in predicting disease outcomes or patient responses to treatment.<\/p>\n<div class=\"btn-container\"><a class=\"btn-sq\" href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/groups\/streptavidin\"><i class=\"fa-solid fa-magnifying-glass\"><\/i>Browse Streptavidin<\/a><br \/>\n<a class=\"btn-sq\" href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/groups\/anti-human-secondary-antibodies\"><i class=\"fa-solid fa-magnifying-glass\"><\/i>Browse Anti-Human Antibodies<\/a><br \/>\n<a class=\"btn-sq\" href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/groups\/fab2\"><i class=\"fa-solid fa-magnifying-glass\"><\/i>Browse F(a<span style=\"font-weight: 400;\">\u2032<\/span>b)<sub>2<\/sub><\/a><\/div>\n<hr \/>\n<h4>References:<\/h4>\n<ul class=\"styled-list\">\n<li><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/25785953\/\">Khalifian, S., Raimondi, G., &amp; Brandacher, G. (2015). The use of luminex assays to measure cytokines.\u00a0<i>The Journal of investigative dermatology<\/i>,\u00a0<i>135<\/i>(4), 1\u20135. https:\/\/doi.org\/10.1038\/jid.2015.36\/<\/a><\/li>\n<li><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/34654825\/\">Egorov, A. I., Converse, R., Griffin, S. M., Bonasso, R., Wickersham, L., Klein, E., Kobylanski, J., Ritter, R., Styles, J. N., Ward, H., Sams, E., Hudgens, E., Dufour, A., &amp; Wade, T. J. (2021). Recreational water exposure and waterborne infections in a prospective salivary antibody study at a Lake Michigan beach.\u00a0<i>Scientific reports<\/i>,\u00a0<i>11<\/i>(1), 20540. https:\/\/doi.org\/10.1038\/s41598-021-00059-2\/<\/a><\/li>\n<li><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/37251407\/\">Lundin, S. B., Kann, H., Fulurija, A., Andersson, B., Nakka, S. S., Andersson, L. M., Gissl\u00e9n, M., &amp; Harandi, A. M. (2023). A novel precision-serology assay for SARS-CoV-2 infection based on linear B-cell epitopes of Spike protein.\u00a0<i>Frontiers in immunology<\/i>,\u00a0<i>14<\/i>, 1166924. https:\/\/doi.org\/10.3389\/fimmu.2023.1166924\/<\/a><\/li>\n<li><a href=\"https:\/\/www.sciencedirect.com\/science\/article\/pii\/S2472630323000535\">Antonia Katharina Hefermehl, Sanne Maria Mathias Hensen, Carina Versantvoort, Andr\u00e9e Rothermel, U\u011fur \u015eahin, Automated glycan-bead coupling for high throughput, highly reproducible anti-glycan antibody analysis, SLAS Technology,2023,,ISSN 2472-6303, https:\/\/doi.org\/10.1016\/j.slast.2023.08.003.5<\/a><\/li>\n<\/ul>\n<div class=\"footnote\">\n<ul>\n<li>Luminex\u00ae and xMAP INTELLIFLEX\u00ae are\u00a0<b>registered trademarks of Luminex Corporation<\/b>.<\/li>\n<\/ul>\n<\/div>\n<p><a href=\"https:\/\/www.jacksonimmuno.com\/technical\/products\/AffiniPure-VHH-secondary-antibodies\"><img decoding=\"async\" loading=\"lazy\" class=\"aligncenter wp-image-9850 size-full\" style=\"width: 100%;\" src=\"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-content\/uploads\/Nano-Secondaries-Advert-ML-GIFFING-4.gif\" alt=\"\" width=\"600\" height=\"100\" \/><\/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<!-- 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 According to Luminex\u00ae (a DiaSorin company), xMAP\u00ae is the world\u2032s most used multiplexing technology, \u00a0a claim that is backed by over 70,000 peer-reviewed publications. These include multiple references to serology assays developed using Jackson ImmunoResearch secondary antibodies, which serve as essential detection reagents for many diagnostic assays using this platform. Principles of the [&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":9697,"comment_status":"closed","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"content-type":""},"categories":[3,20],"tags":[],"acf":[],"_links":{"self":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/10546"}],"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=10546"}],"version-history":[{"count":26,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/10546\/revisions"}],"predecessor-version":[{"id":10634,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/posts\/10546\/revisions\/10634"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/media\/9697"}],"wp:attachment":[{"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/media?parent=10546"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/categories?post=10546"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/www.jacksonimmuno.com\/secondary-antibody-resource\/wp-json\/wp\/v2\/tags?post=10546"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}