{"id":4976,"date":"2025-09-11T16:31:13","date_gmt":"2025-09-11T16:31:13","guid":{"rendered":"https:\/\/www.thermofisher.com\/blog\/life-in-the-lab\/?p=4976"},"modified":"2025-09-11T16:31:16","modified_gmt":"2025-09-11T16:31:16","slug":"guide-to-multiplex-immunoassays-multiplex-elisa-and-more","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/life-in-the-lab\/guide-to-multiplex-immunoassays-multiplex-elisa-and-more\/","title":{"rendered":"Guide to Multiplex Immunoassays: Multiplex ELISA and More"},"content":{"rendered":"\n<figure class=\"wp-block-image aligncenter size-full\"><img loading=\"lazy\" decoding=\"async\" width=\"1200\" height=\"600\" src=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/webinar_profile_image_3d973fdd1fe561b0bdeba5d18c4cea553cd98f07_9948.png\" alt=\"Multiplex Immunoassays: Multiplex ELISA and More\" class=\"wp-image-4978\" srcset=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/webinar_profile_image_3d973fdd1fe561b0bdeba5d18c4cea553cd98f07_9948.png 1200w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/webinar_profile_image_3d973fdd1fe561b0bdeba5d18c4cea553cd98f07_9948-300x150.png 300w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/webinar_profile_image_3d973fdd1fe561b0bdeba5d18c4cea553cd98f07_9948-1024x512.png 1024w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/webinar_profile_image_3d973fdd1fe561b0bdeba5d18c4cea553cd98f07_9948-768x384.png 768w\" sizes=\"auto, (max-width: 1200px) 100vw, 1200px\" \/><\/figure>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<p>Multiplex immunoassays, including multiplex ELISA, are powerful tools for researchers seeking to analyze multiple biomarkers simultaneously.<\/p>\n\n\n\n<p>This guide explores the principles, applications, and advantages of multiplex ELISA and other multiplex immunoassay techniques.<\/p>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<div class=\"wp-block-yoast-seo-table-of-contents yoast-table-of-contents\"><h2>Table of contents<br><\/h2><ul><li><a href=\"#h-what-are-multiplex-immunoassays\" data-level=\"2\">What are multiplex immunoassays?<\/a><ul><li><a href=\"#h-common-research-applications-of-multiplex-immunoassays\" data-level=\"3\">Common research applications of multiplex immunoassays<\/a><\/li><\/ul><\/li><li><a href=\"#h-what-are-the-different-types-of-immunoassays\" data-level=\"2\">What are the different types of immunoassays?<\/a><ul><li><a href=\"#h-electrochemiluminescence-immunoassay-eclia\" data-level=\"3\">Electrochemiluminescence Immunoassay (ECLIA)<\/a><\/li><li><a href=\"#h-olink-pea\" data-level=\"3\">Olink PEA<\/a><\/li><li><a href=\"#h-luminex-bead-based\" data-level=\"3\">Luminex \/ Bead-Based<\/a><\/li><\/ul><\/li><li><a href=\"#h-when-to-use-a-multiplex-elisa\" data-level=\"2\">When to use a multiplex ELISA<\/a><\/li><li><a href=\"#h-multiplex-immunoassay-vs-elisa-similarities-and-differences\" data-level=\"2\">Multiplex immunoassay vs. ELISA: similarities and differences<\/a><ul><li><a href=\"#h-quality-of-results-and-reproducibility\" data-level=\"3\">Quality of results and reproducibility<\/a><\/li><li><a href=\"#h-difficulty-and-time-to-perform\" data-level=\"3\">Difficulty and time to perform<\/a><\/li><li><a href=\"#h-cost\" data-level=\"3\">Cost<\/a><\/li><\/ul><\/li><li><a href=\"#h-summary-of-the-benefits-of-multiplexing-immunoassays\" data-level=\"2\">Summary of the benefits of multiplexing immunoassays<\/a><\/li><\/ul><\/div>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"h-what-are-multiplex-immunoassays\">What are multiplex immunoassays? <\/h2>\n\n\n\n<p>Multiplex immunoassays are advanced techniques used in biomedical research to simultaneously measure multiple analytes (e.g., proteins, peptides, or antibodies) in a single well with a small sample volume (e.g. 1-50 \u00b5L) of plasma, serum, cell culture supernatants, or other bodily \ufb02uids. By allowing the detection of several biomarkers at once, these assays help provide a more comprehensive overview of biological processes and disease states compared to traditional single-analyte assays and are especially valuable when sample volume is limited.<\/p>\n\n\n\n<p><a href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/life-science\/antibodies\/immunoassays\/elisa-kits.html\" target=\"_blank\" rel=\"noreferrer noopener\">Enzyme-Linked Immunosorbent Assay (ELISA)<\/a> is a common and widely used method to perform sample analysis and can accurately detect and quantify individual proteins with high specificity and sensitivity. The selectivity of ELISA is achieved using qualified single- or double-antibody sandwich technology, and accurate quantitation using calibrated standards. ELISAs are performed in a 96-well format with results typically ready after ~4 hours on a <a href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/life-science\/lab-equipment\/microplate-instruments\/plate-readers\/models.html\" target=\"_blank\" rel=\"noreferrer noopener\">microplate reader.<\/a> Results obtained with ELISAs are generally reproducible and consistent with biologically relevant sensitivity levels and dynamic ranges.<\/p>\n\n\n\n<p>While ELISA is a widely trusted method of protein analysis in general, multiplexing methods enable the measurement of multiple analytes simultaneously in a single well requiring smaller sample input and less hands-on time than ELISA, helping reduce time to results, cost and labor.<\/p>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h3 class=\"wp-block-heading\" id=\"h-common-research-applications-of-multiplex-immunoassays\">Common research applications of multiplex immunoassays<\/h3>\n\n\n\n<p>Multiplex immunoassays are widely utilized in various research fields, including:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Biomarker Discovery<\/strong>: Identifying new biomarkers for diseases.<\/li>\n\n\n\n<li><strong>Immune Response Studies<\/strong>: Understanding how the immune system responds to infections or treatments.<\/li>\n\n\n\n<li><strong>Drug and Vaccine development:<\/strong> Measuring inflammatory and cytokine responses pre- and post-treatment<\/li>\n\n\n\n<li><strong>Clinical Diagnostics<\/strong>: Diagnosing and monitoring diseases.<\/li>\n\n\n\n<li><strong>Pharmacodynamics and Toxicology Studies<\/strong>: Assessing the effects of drugs and their safety profiles.<\/li>\n<\/ul>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"h-what-are-the-different-types-of-immunoassays\">What are the different types of immunoassays? <\/h2>\n\n\n\n<p>There are several types of multiplex immunoassays available, and three are described below.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\" id=\"h-electrochemiluminescence-immunoassay-eclia\">Electrochemiluminescence Immunoassay (ECLIA)<\/h3>\n\n\n\n<p>Electrochemiluminescence (ECL) assays use electrochemical and chemiluminescent principles to detect multiple analytes. This method offers high sensitivity and a wide dynamic range, making it suitable for applications requiring precise quantification of low-abundance biomarkers.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\" id=\"h-olink-pea\">Olink PEA<\/h3>\n\n\n\n<p>The <a href=\"https:\/\/olink.com\/technology\/what-is-pea\" target=\"_blank\" rel=\"noreferrer noopener\">Olink Proximity Extension Assay (PEA)<\/a> is a highly specific and sensitive technology that uses DNA-labeled antibody pairs to detect proteins. When the antibodies bind to their target, their DNA tags come into proximity and are extended, allowing for subsequent quantification using qPCR or next-generation sequencing (NGS). Olink PEA allows high multiplexing of up to 5,000+ proteins with high-specificity and high-sensitivity.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\" id=\"h-luminex-bead-based\">Luminex \/ Bead-Based<\/h3>\n\n\n\n<p><a href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/life-science\/antibodies\/immunoassays\/procartaplex-assays-luminex\/features.html#xmap-technology\" target=\"_blank\" rel=\"noreferrer noopener\">Luminex\u00ae xMAP\u00ae (multi-analyte profiling) technology<\/a> uses color-coded beads, which are dyed with different concentration of fluorophores to generate bead sets that can be easily differentiated (Figure 1A). Individual bead sets are coated with specific antibodies to capture either one target analyte or two different targets on the same bead as used in Dual Reporter assays.<\/p>\n\n\n\n<p>The captured analyte from a sample is detected using an analyte-specific biotinylated antibody that binds to the appropriate epitope of the immobilized analyte, plus streptavidin-conjugated R-phycoerythrin (S-RPE) or streptavidin-BV421 for Dual Reporter assay.<\/p>\n\n\n\n<p>For detection of the immunoassay sandwich complex, <a href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/life-science\/antibodies\/immunoassays\/procartaplex-assays-luminex\/luminex-instruments.html\" target=\"_blank\" rel=\"noreferrer noopener\">Luminex\u2122 instruments<\/a> use either light-emitting diodes (LEDs) for excitation of each fluorescent bead combined with a CCD camera for bead and analyte detection (MagPix, discontinued), or a flow-based detection system using with a red and green laser for excitation of each fluorescent bead and measurement of the reporter dye (Luminex 200, FLEXMAP 3D, INTELLIFLEX, INTELLIFLEX DR-SE) and additional violet laser for measuring the second reporter in Dual Reporter assays (only available in INTELLIFLEX DR-SE).<\/p>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<figure data-wp-context=\"{&quot;imageId&quot;:&quot;6ab500a242418&quot;}\" data-wp-interactive=\"core\/image\" data-wp-key=\"6ab500a242418\" class=\"wp-block-image aligncenter size-large wp-lightbox-container\"><img loading=\"lazy\" decoding=\"async\" width=\"1024\" height=\"271\" data-wp-class--hide=\"state.isContentHidden\" data-wp-class--show=\"state.isContentVisible\" data-wp-init=\"callbacks.setButtonStyles\" data-wp-on--click=\"actions.showLightbox\" data-wp-on--load=\"callbacks.setButtonStyles\" data-wp-on-window--resize=\"callbacks.setButtonStyles\" src=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA2-1024x271.jpg\" alt=\"Figure 1. How Luminex xMAP technology works. (A) Capture antibodies are bound to distinct Luminex beads which are internally dyed, and this allows the analysis of multiple analytes in a single well. (B) Samples mixed with bead sets bind analytes of interest via capture antibodies, and fluorescently labeled detection antibodies form an antibody-antigen sandwich. (C) Completed assays are read on a Luminex instrument, where one laser identifies the bead type and analyte, and another laser measures the magnitude of the bound analyte via PE-derived signal. \" class=\"wp-image-4979\" srcset=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA2-1024x271.jpg 1024w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA2-300x79.jpg 300w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA2-768x203.jpg 768w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA2.jpg 1453w\" sizes=\"auto, (max-width: 1024px) 100vw, 1024px\" \/><button\n\t\t\tclass=\"lightbox-trigger\"\n\t\t\ttype=\"button\"\n\t\t\taria-haspopup=\"dialog\"\n\t\t\taria-label=\"Enlarge\"\n\t\t\tdata-wp-init=\"callbacks.initTriggerButton\"\n\t\t\tdata-wp-on--click=\"actions.showLightbox\"\n\t\t\tdata-wp-style--right=\"state.imageButtonRight\"\n\t\t\tdata-wp-style--top=\"state.imageButtonTop\"\n\t\t>\n\t\t\t<svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"12\" height=\"12\" fill=\"none\" viewBox=\"0 0 12 12\">\n\t\t\t\t<path fill=\"#fff\" d=\"M2 0a2 2 0 0 0-2 2v2h1.5V2a.5.5 0 0 1 .5-.5h2V0H2Zm2 10.5H2a.5.5 0 0 1-.5-.5V8H0v2a2 2 0 0 0 2 2h2v-1.5ZM8 12v-1.5h2a.5.5 0 0 0 .5-.5V8H12v2a2 2 0 0 1-2 2H8Zm2-12a2 2 0 0 1 2 2v2h-1.5V2a.5.5 0 0 0-.5-.5H8V0h2Z\" \/>\n\t\t\t<\/svg>\n\t\t<\/button><figcaption class=\"wp-element-caption\">Figure 1.<strong> How Luminex xMAP technology works.<\/strong> (A) Capture antibodies are bound to distinct Luminex beads which are internally dyed, and this allows the analysis of multiple analytes in a single well. (B) Samples mixed with bead sets bind analytes of interest via capture antibodies, and fluorescently labeled detection antibodies form an antibody-antigen sandwich. (C) Completed assays are read on a Luminex instrument, where one laser identifies the bead type and analyte, and another laser measures the magnitude of the bound analyte via PE-derived signal.<\/figcaption><\/figure>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<p>This Luminex technology is versatile and can be used for detecting both proteins and nucleic acids, making it a <a href=\"https:\/\/www.thermofisher.com\/blog\/behindthebench\/two-applications-one-instrument-assay\/\" target=\"_blank\" rel=\"noreferrer noopener\">dual-capability platform<\/a> for a wide range of applications in research. As each coated bead is individually identifiable for a specific analyte, multiple beads can be combined to simultaneously measure the levels of up to 500 targets for nucleic acid and typically no more than 80 targets for proteins due to biological interference in a single sample. Therefore, this bead-based approach allows for high-throughput and flexible multiplexing.<\/p>\n\n\n\n<div style=\"height:25px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<div class=\"wp-block-buttons is-content-justification-center is-layout-flex wp-container-core-buttons-is-layout-16018d1d wp-block-buttons-is-layout-flex\">\n<div class=\"wp-block-button has-custom-width wp-block-button__width-75 is-style-fill\"><a class=\"wp-block-button__link has-white-color has-text-color has-background has-link-color has-medium-font-size has-text-align-center has-custom-font-size wp-element-button\" href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/global\/forms\/magpix-system-demo.html\" style=\"border-radius:0px;background-color:#ee3134\" target=\"_blank\" rel=\"noreferrer noopener\"><strong>Request a Luminex technology demo<\/strong><\/a><\/div>\n<\/div>\n\n\n\n<div style=\"height:25px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"h-when-to-use-a-multiplex-elisa\">When to use a multiplex ELISA<\/h2>\n\n\n\n<p>While ELISA is a trusted method of protein analysis, multiplexing methods that enable the measurement of multiple analytes simultaneously in a single well address a number of specific limitations:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>ELISA allows for the measurement of only one analyte at a time in a given sample, limiting investigators\u2019 increasing need to measure multiple targets for a holistic biological understanding of protein interactions.<\/li>\n\n\n\n<li>The limited sample volume available may limit the number of times analyses can be conducted. This is especially true in small animal research, in pediatric testing, and in microplate assays providing limited sample volume. The ability to assay multiple analytes in a single small-volume sample enables more effective use of each sample.<\/li>\n\n\n\n<li>Difficulties in data interpretation can arise when comparing analyte levels measured by multiple ELISAs, each assay having been performed with different sample aliquots and each subject to systematic errors leading to decreased precision and accuracy.<\/li>\n\n\n\n<li>Many analytes require assays with broad dynamic ranges to avoid repeat testing or out-of-range values. Multiplex assays can be designed to have large dynamic ranges for all of the analytes, or customer ranges tailored to various expected analyte concentrations.<\/li>\n<\/ul>\n\n\n\n<p>The analysis of selected proteins in, for example, serum, plasma, cell lysates or urine is key to the study of inflammation, metabolism, cell signaling, cancer, cardiovascular disease, toxicology, neuroscience, and many other research and clinical areas. The ever-growing list of cytokines, chemokines, and other growth and differentiation factors, as well as their complex interactive networks in health and disease, has made it crucial to evaluate them in relevant groups rather than individually. Often, for example, the levels of cytokines or chemokines within a pathway in relation to each other are far more relevant than their presence or absence, or than the absolute levels of individual proteins.<\/p>\n\n\n\n<p>Luminex\u2122 xMAP\u2122 assays have increasingly been adopted for sample analysis, and over 98,000 scientific publications are available demonstrating high precision and reproducible results. These studies have covered a broad range of common applications, including:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li>Measurement of soluble analytes in serum, plasma, urine, cerebral spinal fluid (CSF), lavage fluids, cell culture supernatants and other sample types from human, monkey, mouse, rat, swine, canine and other species.<\/li>\n\n\n\n<li>Basic research\u2014<em>in vitro<\/em> and <em>in vivo<\/em> studies<\/li>\n\n\n\n<li>Preclinical studies\u2014<em>in vitro<\/em> and <em>in vivo<\/em> models<\/li>\n\n\n\n<li>Screening batches of sera for the presence\/absence of defined markers or mediators<\/li>\n\n\n\n<li>Quantitative confirmation of data for targets identified from proteomic or genomic analysis<\/li>\n<\/ul>\n\n\n\n<p><a href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/life-science\/antibodies\/immunoassays\/procartaplex-assays-luminex\/procartaplex-immunoassays.html\" target=\"_blank\" rel=\"noreferrer noopener\">Invitrogen\u2122 ProcartaPlex\u2122 multiplex immunoassays\u00a0<\/a>are based on this Luminex xMAP technology, and provide a versatile platform that gives users more flexibility and a greater array of options for analyte detection.<\/p>\n\n\n\n<p>Whether you are testing for single or multiple analytes, ProcartaPlex multiplex assays deliver accurate analytical performance using efficient, easy-to-follow protocols. These assays are developed by ELISA experts and undergo rigorous development, validation, manufacturing, and quality control processes comparable to our Invitrogen ELISAs.<\/p>\n\n\n\n<p>In addition, multiplex assays undergo additional testing to ensure that there is no crosstalk between the assays. &nbsp;Each ProcartaPlex multiplex assay and ELISA assay is fully qualified with the sample type stated&nbsp; (i.e., species-specific serum, plasma, and cell culture supernatants), and each lot is evaluated based on the following performance characteristics:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Specificity<\/strong>\u2014each antibody pair is thoroughly tested in development to help ensure it detects the correct analyte and does not cross-react with other analytes<\/li>\n\n\n\n<li><strong>Sensitivity<\/strong>\u2014each analyte is evaluated for both functional sensitivity (differentiation from background) and lower limit of detection (LLOD)<\/li>\n\n\n\n<li><strong>Precision\/accuracy<\/strong>\u2014multiplex assays have good intra-assay precision (&lt;15% CV), inter-assay precision (&lt;15% CV), and lot-to-lot consistency (&lt;30% CV)<\/li>\n<\/ul>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<figure data-wp-context=\"{&quot;imageId&quot;:&quot;6ab500a2431bb&quot;}\" data-wp-interactive=\"core\/image\" data-wp-key=\"6ab500a2431bb\" class=\"wp-block-image aligncenter size-full wp-lightbox-container\"><img loading=\"lazy\" decoding=\"async\" width=\"983\" height=\"388\" data-wp-class--hide=\"state.isContentHidden\" data-wp-class--show=\"state.isContentVisible\" data-wp-init=\"callbacks.setButtonStyles\" data-wp-on--click=\"actions.showLightbox\" data-wp-on--load=\"callbacks.setButtonStyles\" data-wp-on-window--resize=\"callbacks.setButtonStyles\" src=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA3.jpg\" alt=\"Figure 2. Dynamic range and correlation to ELISA. (A) Standard curves were generated for human IL-1\u00df using an Invitrogen ProcartaPlex multiplex assay and Invitrogen ELISA. The dynamic range of the multiplex assay is much larger than the ELISA. (B) Murine GM-CSF in cell culture supernatant was tested by ELISA (y-axis) and in an Invitrogen multiplex assay (x-axis). Correlation of values over 3 logs of sample dilution was 0.9868.\" class=\"wp-image-4980\" srcset=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA3.jpg 983w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA3-300x118.jpg 300w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA3-768x303.jpg 768w\" sizes=\"auto, (max-width: 983px) 100vw, 983px\" \/><button\n\t\t\tclass=\"lightbox-trigger\"\n\t\t\ttype=\"button\"\n\t\t\taria-haspopup=\"dialog\"\n\t\t\taria-label=\"Enlarge\"\n\t\t\tdata-wp-init=\"callbacks.initTriggerButton\"\n\t\t\tdata-wp-on--click=\"actions.showLightbox\"\n\t\t\tdata-wp-style--right=\"state.imageButtonRight\"\n\t\t\tdata-wp-style--top=\"state.imageButtonTop\"\n\t\t>\n\t\t\t<svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"12\" height=\"12\" fill=\"none\" viewBox=\"0 0 12 12\">\n\t\t\t\t<path fill=\"#fff\" d=\"M2 0a2 2 0 0 0-2 2v2h1.5V2a.5.5 0 0 1 .5-.5h2V0H2Zm2 10.5H2a.5.5 0 0 1-.5-.5V8H0v2a2 2 0 0 0 2 2h2v-1.5ZM8 12v-1.5h2a.5.5 0 0 0 .5-.5V8H12v2a2 2 0 0 1-2 2H8Zm2-12a2 2 0 0 1 2 2v2h-1.5V2a.5.5 0 0 0-.5-.5H8V0h2Z\" \/>\n\t\t\t<\/svg>\n\t\t<\/button><figcaption class=\"wp-element-caption\">Figure 2. <strong>Dynamic range and correlation to ELISA.\u00a0<\/strong>(A) Standard curves were generated for human IL-1\u00df using an Invitrogen ProcartaPlex multiplex assay and Invitrogen ELISA. The dynamic range of the multiplex assay is much larger than the ELISA. (B) Murine GM-CSF in cell culture supernatant was tested by ELISA (y-axis) and in an Invitrogen multiplex assay (x-axis). Correlation of values over 3 logs of sample dilution was 0.9868.<\/figcaption><\/figure>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<p>To demonstrate the power of multiplexing, 30 samples were analyzed: 23 untreated human serum samples and 7 plasma samples treated with 100ng\/ml LPS for 20 hours . Expression pattern of low expressed targets are shown in the figure below. Data were generated using the <a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/EPX800-10080-901\" target=\"_blank\" rel=\"noreferrer noopener\">Invitrogen\u2122 ProcartaPlex\u2122 Human Immune Response Panel, 80plex.<\/a><\/p>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<figure data-wp-context=\"{&quot;imageId&quot;:&quot;6ab500a2435d2&quot;}\" data-wp-interactive=\"core\/image\" data-wp-key=\"6ab500a2435d2\" class=\"wp-block-image aligncenter size-full wp-lightbox-container\"><img loading=\"lazy\" decoding=\"async\" width=\"1025\" height=\"363\" data-wp-class--hide=\"state.isContentHidden\" data-wp-class--show=\"state.isContentVisible\" data-wp-init=\"callbacks.setButtonStyles\" data-wp-on--click=\"actions.showLightbox\" data-wp-on--load=\"callbacks.setButtonStyles\" data-wp-on-window--resize=\"callbacks.setButtonStyles\" src=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA4.jpg\" alt=\"\" class=\"wp-image-4981\" srcset=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA4.jpg 1025w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA4-300x106.jpg 300w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA4-768x272.jpg 768w\" sizes=\"auto, (max-width: 1025px) 100vw, 1025px\" \/><button\n\t\t\tclass=\"lightbox-trigger\"\n\t\t\ttype=\"button\"\n\t\t\taria-haspopup=\"dialog\"\n\t\t\taria-label=\"Enlarge\"\n\t\t\tdata-wp-init=\"callbacks.initTriggerButton\"\n\t\t\tdata-wp-on--click=\"actions.showLightbox\"\n\t\t\tdata-wp-style--right=\"state.imageButtonRight\"\n\t\t\tdata-wp-style--top=\"state.imageButtonTop\"\n\t\t>\n\t\t\t<svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"12\" height=\"12\" fill=\"none\" viewBox=\"0 0 12 12\">\n\t\t\t\t<path fill=\"#fff\" d=\"M2 0a2 2 0 0 0-2 2v2h1.5V2a.5.5 0 0 1 .5-.5h2V0H2Zm2 10.5H2a.5.5 0 0 1-.5-.5V8H0v2a2 2 0 0 0 2 2h2v-1.5ZM8 12v-1.5h2a.5.5 0 0 0 .5-.5V8H12v2a2 2 0 0 1-2 2H8Zm2-12a2 2 0 0 1 2 2v2h-1.5V2a.5.5 0 0 0-.5-.5H8V0h2Z\" \/>\n\t\t\t<\/svg>\n\t\t<\/button><figcaption class=\"wp-element-caption\">Figure 3. <strong>Differences in the expression pattern of 80 targets in untreated human serum samples and pre-treated plasma samples were measured with <a href=\"https:\/\/www.thermofisher.com\/order\/catalog\/product\/EPX800-10080-901\" target=\"_blank\" rel=\"noreferrer noopener\">Invitrogen\u2122 ProcartaPlex\u2122 Human Immune Response Panel, 80plex.<\/a><\/strong> A selection of low expressed analytes are shown here. Analytes are displayed on the x-axis, and mean concentration for untreated (blue lines) versus treated (red lines) samples for each analyte on the y-axis.<\/figcaption><\/figure>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"h-multiplex-immunoassay-vs-elisa-similarities-and-differences\">Multiplex immunoassay vs. ELISA: similarities and differences<\/h2>\n\n\n\n<h3 class=\"wp-block-heading\" id=\"h-quality-of-results-and-reproducibility\">Quality of results and reproducibility<\/h3>\n\n\n\n<p>Both multiplex immunoassays and traditional ELISA offer high-quality results. However, multiplex immunoassays can help provide more comprehensive data as they measure multiple analytes simultaneously, leading to better insights into complex biological systems. Reproducibility can be maintained across different runs, provided that the assays are well-validated.<\/p>\n\n\n\n<h3 class=\"wp-block-heading\" id=\"h-difficulty-and-time-to-perform\">Difficulty and time to perform<\/h3>\n\n\n\n<p>Multiplex immunoassays can be more complex to set up and optimize compared to single-analyte ELISA. However, once established, they save significant time by allowing the simultaneous measurement of multiple targets. This reduces the number of assays needed and speeds up data generation.<\/p>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<figure data-wp-context=\"{&quot;imageId&quot;:&quot;6ab500a243abe&quot;}\" data-wp-interactive=\"core\/image\" data-wp-key=\"6ab500a243abe\" class=\"wp-block-image aligncenter size-full wp-lightbox-container\"><img loading=\"lazy\" decoding=\"async\" width=\"681\" height=\"340\" data-wp-class--hide=\"state.isContentHidden\" data-wp-class--show=\"state.isContentVisible\" data-wp-init=\"callbacks.setButtonStyles\" data-wp-on--click=\"actions.showLightbox\" data-wp-on--load=\"callbacks.setButtonStyles\" data-wp-on-window--resize=\"callbacks.setButtonStyles\" src=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA5.png\" alt=\"Figure 4. Time savings for Invitrogen ProcartaPlex multiplex assay kits vs. ELISAs. The time savings of multiplexing becomes more significant as the number of analytes increases.\" class=\"wp-image-4982\" srcset=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA5.png 681w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA5-300x150.png 300w\" sizes=\"auto, (max-width: 681px) 100vw, 681px\" \/><button\n\t\t\tclass=\"lightbox-trigger\"\n\t\t\ttype=\"button\"\n\t\t\taria-haspopup=\"dialog\"\n\t\t\taria-label=\"Enlarge\"\n\t\t\tdata-wp-init=\"callbacks.initTriggerButton\"\n\t\t\tdata-wp-on--click=\"actions.showLightbox\"\n\t\t\tdata-wp-style--right=\"state.imageButtonRight\"\n\t\t\tdata-wp-style--top=\"state.imageButtonTop\"\n\t\t>\n\t\t\t<svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"12\" height=\"12\" fill=\"none\" viewBox=\"0 0 12 12\">\n\t\t\t\t<path fill=\"#fff\" d=\"M2 0a2 2 0 0 0-2 2v2h1.5V2a.5.5 0 0 1 .5-.5h2V0H2Zm2 10.5H2a.5.5 0 0 1-.5-.5V8H0v2a2 2 0 0 0 2 2h2v-1.5ZM8 12v-1.5h2a.5.5 0 0 0 .5-.5V8H12v2a2 2 0 0 1-2 2H8Zm2-12a2 2 0 0 1 2 2v2h-1.5V2a.5.5 0 0 0-.5-.5H8V0h2Z\" \/>\n\t\t\t<\/svg>\n\t\t<\/button><figcaption class=\"wp-element-caption\">Figure 4. <strong>Time savings for Invitrogen ProcartaPlex multiplex assay kits vs. ELISAs<\/strong>. The time savings of multiplexing becomes more significant as the number of analytes increases.<\/figcaption><\/figure>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h3 class=\"wp-block-heading\" id=\"h-cost\">Cost<\/h3>\n\n\n\n<p>While the initial setup and reagents for multiplex immunoassays can be more expensive than single-analyte ELISA, the overall cost per analyte is often lower due to the reduced number of assays and samples required. The cost-effectiveness increases with the number of analytes measured.<\/p>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<figure data-wp-context=\"{&quot;imageId&quot;:&quot;6ab500a243e8a&quot;}\" data-wp-interactive=\"core\/image\" data-wp-key=\"6ab500a243e8a\" class=\"wp-block-image aligncenter size-full wp-lightbox-container\"><img loading=\"lazy\" decoding=\"async\" width=\"684\" height=\"715\" data-wp-class--hide=\"state.isContentHidden\" data-wp-class--show=\"state.isContentVisible\" data-wp-init=\"callbacks.setButtonStyles\" data-wp-on--click=\"actions.showLightbox\" data-wp-on--load=\"callbacks.setButtonStyles\" data-wp-on-window--resize=\"callbacks.setButtonStyles\" src=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA6.png\" alt=\"\" class=\"wp-image-4983\" srcset=\"https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA6.png 684w, https:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA6-287x300.png 287w\" sizes=\"auto, (max-width: 684px) 100vw, 684px\" \/><button\n\t\t\tclass=\"lightbox-trigger\"\n\t\t\ttype=\"button\"\n\t\t\taria-haspopup=\"dialog\"\n\t\t\taria-label=\"Enlarge\"\n\t\t\tdata-wp-init=\"callbacks.initTriggerButton\"\n\t\t\tdata-wp-on--click=\"actions.showLightbox\"\n\t\t\tdata-wp-style--right=\"state.imageButtonRight\"\n\t\t\tdata-wp-style--top=\"state.imageButtonTop\"\n\t\t>\n\t\t\t<svg xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"12\" height=\"12\" fill=\"none\" viewBox=\"0 0 12 12\">\n\t\t\t\t<path fill=\"#fff\" d=\"M2 0a2 2 0 0 0-2 2v2h1.5V2a.5.5 0 0 1 .5-.5h2V0H2Zm2 10.5H2a.5.5 0 0 1-.5-.5V8H0v2a2 2 0 0 0 2 2h2v-1.5ZM8 12v-1.5h2a.5.5 0 0 0 .5-.5V8H12v2a2 2 0 0 1-2 2H8Zm2-12a2 2 0 0 1 2 2v2h-1.5V2a.5.5 0 0 0-.5-.5H8V0h2Z\" \/>\n\t\t\t<\/svg>\n\t\t<\/button><figcaption class=\"wp-element-caption\">Figure 5. <strong>Cost and time savings for Invitrogen ProcartaPlex multiplex assay vs. ELISA kits<\/strong>. The x-axis shows the number of analytes tested, and the y-axis the total assay cost. The cost advantage of Invitrogen multiplexing assays becomes more and more significant as the number of analytes increases.<\/figcaption><\/figure>\n\n\n\n<div style=\"height:25px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<div class=\"wp-block-buttons is-content-justification-center is-layout-flex wp-container-core-buttons-is-layout-16018d1d wp-block-buttons-is-layout-flex\">\n<div class=\"wp-block-button has-custom-width wp-block-button__width-75 is-style-fill\"><a class=\"wp-block-button__link has-white-color has-text-color has-background has-link-color has-medium-font-size has-text-align-center has-custom-font-size wp-element-button\" href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/global\/forms\/luminex-instrument-quote-request.html\" style=\"border-radius:0px;background-color:#ee3134\" target=\"_blank\" rel=\"noreferrer noopener\"><strong>Get a custom quote for multiple targets with Luminex<\/strong><\/a><\/div>\n<\/div>\n\n\n\n<div style=\"height:25px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h2 class=\"wp-block-heading\" id=\"h-summary-of-the-benefits-of-multiplexing-immunoassays\">Summary of the benefits of multiplexing immunoassays<\/h2>\n\n\n\n<p>The Luminex multiplex bead-based ProcartaPlex immunoassays have a number of significant advantages compared with traditional ELISA technology, including:<\/p>\n\n\n\n<ul class=\"wp-block-list\">\n<li><strong>Time savings<\/strong>\u2014ProcartaPlex assays allow testing of up to 80 analytes in a single sample. In the same time it takes to set up one ELISA, multiple analytes can be measured, significantly reducing labor time.<\/li>\n\n\n\n<li><strong>Smaller sample volume<\/strong>\u2014depending on the expression levels, multiplex assays require 25-50 \u00b5L of sample or less for 384-well format, while still obtaining accurate results for all analytes.<\/li>\n\n\n\n<li><strong>Broad dynamic range<\/strong>\u2014provides the ability to reliably detect proteins across a broad concentration range.<\/li>\n\n\n\n<li><strong>High throughput<\/strong>\u2014the Luminex system reads from conventional 96-well and 384-well microtiter plate. Combined with the ability to read up to 80analytes per sample, this provides a high-throughput path to data collection.<\/li>\n<\/ul>\n\n\n\n<p>Field Application Specialists (FAS) are available to educate you on multiplexing, answer questions, and help support all aspects of running and analyzing an Invitrogen multiplex assay. For any questions, please contact us at <a href=\"mailto:LuminexFAS@thermofisher.com\" target=\"_blank\" rel=\"noreferrer noopener nofollow\">LuminexFAS@thermofisher.com<\/a> or access the <a href=\"https:\/\/www.thermofisher.com\/at\/en\/home\/global\/forms\/life-science\/multiplex-academy.html\" target=\"_blank\" rel=\"noreferrer noopener\">Multiplex Academy<\/a> for a self-learning platform.<\/p>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<h2 class=\"wp-block-heading\">Learn more about the product in this blog:<\/h2>\n\n\n\n<h3 class=\"wp-block-heading\"><strong>Resources and Solutions for Luminex and ProcartaPlex immunoassay<\/strong><\/h3>\n\n\n\n<ul class=\"wp-block-list\">\n<li><a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/life-science\/antibodies\/immunoassays\/procartaplex-assays-luminex.html?icid=bid_pca_aup_ro1_co_1434_pjt9046_COL020405_0s0_blg_op_awa_kt_s00_assay10\" target=\"_blank\" rel=\"noreferrer noopener\">Luminex platform<\/a> \u2013 Quantitative protein and RNA assays with superior throughput<\/li>\n\n\n\n<li><a href=\"https:\/\/assets.thermofisher.com\/TFS-Assets\/BID\/Reference-Materials\/luminex-instrument-assay-guide-selection-guide.pdf\" target=\"_blank\" rel=\"noreferrer noopener\">Luminex instrument and assay guide \u2013 High-Throughput multiplexing of protein and gene expression analysis<\/a><\/li>\n\n\n\n<li><a href=\"https:\/\/www.thermofisher.com\/order\/luminex\/?icid=bid_pca_aup_ro1_co_1434_pjt9046_COL020405_0s0_blg_op_awa_kt_s00_assay11\" target=\"_blank\" rel=\"noreferrer noopener\">ProcartaPlex Panel Configurator<\/a> \u2013 This online tool helps you design a custom multiplex panel for your specific application<\/li>\n\n\n\n<li><a href=\"https:\/\/www.thermofisher.com\/us\/en\/home\/technical-resources\/technical-reference-library\/antibodies-immunoassays-support-center\/luminex-assays-support.html?icid=bid_pca_aup_ro1_co_1434_pjt9046_COL020405_0s0_blg_op_awa_kt_s00_assay13\" target=\"_blank\" rel=\"noreferrer noopener\">ProcartaPlex Assays Support<\/a> \u2013 Explore the \u201cGetting Started\u201d and \u201cTroubleshooting\u201d sections for solutions to top inquiries and common problems.<\/li>\n<\/ul>\n\n\n\n<div style=\"height:15px\" aria-hidden=\"true\" class=\"wp-block-spacer\"><\/div>\n\n\n\n<p><em>##<\/em><\/p>\n\n\n\n<p><em>For Research Use Only. Not for use in diagnostic procedures. <\/em>&nbsp;&nbsp;<\/p>\n\n\n\n<p><em>\u00a9 2025 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.<\/em><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Multiplex immunoassays, including multiplex ELISA, are powerful tools for researchers seeking to analyze multiple biomarkers simultaneously. This guide explores the principles, applications, and advantages of multiplex ELISA and other multiplex immunoassay techniques. What are multiplex immunoassays? Multiplex immunoassays are advanced techniques used in biomedical research to simultaneously measure multiple analytes (e.g., proteins, peptides, or antibodies)<\/p>\n","protected":false},"author":1670,"featured_media":4977,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_kad_blocks_custom_css":"","_kad_blocks_head_custom_js":"","_kad_blocks_body_custom_js":"","_kad_blocks_footer_custom_js":"","_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[976,4],"tags":[],"division":[],"class_list":{"0":"post-4976","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-connect-to-science","8":"category-general","9":"entry"},"_selected_authors":[],"_selected_reviewers":[],"acf":[],"yoast_head":"<!-- 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multiplex ELISA and other multiplex immunoassay techniques.\" \/>\n<meta property=\"og:url\" content=\"https:\/\/www.thermofisher.com\/blog\/life-in-the-lab\/guide-to-multiplex-immunoassays-multiplex-elisa-and-more\/\" \/>\n<meta property=\"og:site_name\" content=\"Life in the Lab\" \/>\n<meta property=\"article:publisher\" content=\"https:\/\/www.facebook.com\/thermofisher\" \/>\n<meta property=\"article:published_time\" content=\"2025-09-11T16:31:13+00:00\" \/>\n<meta property=\"article:modified_time\" content=\"2025-09-11T16:31:16+00:00\" \/>\n<meta property=\"og:image\" content=\"http:\/\/admin.acceleratingscience.com\/life-in-the-lab\/wp-content\/uploads\/sites\/10\/2025\/09\/ELISA1.png\" \/>\n\t<meta property=\"og:image:width\" content=\"468\" \/>\n\t<meta property=\"og:image:height\" content=\"234\" \/>\n\t<meta property=\"og:image:type\" content=\"image\/png\" \/>\n<meta name=\"author\" content=\"Dana D&#039;Amico\" \/>\n<meta name=\"twitter:card\" 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