CINXE.COM

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<img class="img-banner" src="/sites/default/files/STRY0320271_SBD_WU_ACDbio-Sequential_RNA_TA-min.jpg?q=1"><h1 class="general-title">Sequential RNA Protein Detection</h1><h2 class="banner-sub-title-seo">RNAscope™ Applications</h2></div><div class="content-container container newapplication"><div id="node-53409" class="node node-application" about="/science/applications/research-solutions/sequential-rna-protein-detection" typeof="sioc:Item foaf:Document"><div class="nav-tab-container"><ul class="nav nav-tabs page-tabs application-bxslider" id="myTab"><li class=""><a href="/rna-protein-co-detection-imaging-mass-cytometry-and-rnascope-assay">RNAscope HiPlex-IMC™ Co-Detection</a></li><li class=""><a href="/node/425364">Integrated Co-Detection</a></li><li class=" active"><a href="/node/53409">Sequential RNA Protein Detection</a></li></ul></div><div class="tab-content"><div id="tab0" class="tab-pane fade in active profile"><h1 class="top-profile">Dual ISH and IHC</h1><div class="sample-image"><div class="loader-image-gallery"></div><div id="film_roll" class="sample-slider"><div><img src="https://acdbio.com/sites/default/files/styles/film_roll_slider/public/Dual%20ISH-IH-From%20Bradley%20Spencer_%20triple%20lung%20_PPI_CAM5.2_DAPI_1_0.jpg?itok=hZoVrz8x" width="765" height="451" alt="Human lung cancer with human PPIB positive control mRNA probe in red, CAM5.2 acidic cytokeratin immunohistochemical staining in green with DAPI as counterstain."><p class="slider-below-text"> Human lung cancer with human PPIB positive control mRNA probe in red, CAM5.2 acidic cytokeratin immunohistochemical staining in green with DAPI as counterstain.</p></div><div><img src="https://acdbio.com/sites/default/files/styles/film_roll_slider/public/Dual%20ISH-IHC%20-%20Paola%20data_%20OLFM4%20ISH%20and%20Lysozume%20AB%20IHC_0.png?itok=mm2zBtqQ" width="765" height="451" alt="Dual ISH-IHC - Detection of OLFM4 using RNAscope technology and Lyzozyme using IHC"><p class="slider-below-text"> Dual ISH-IHC - Detection of OLFM4 using RNAscope technology and Lyzozyme using IHC</p></div><div><img src="https://acdbio.com/sites/default/files/styles/film_roll_slider/public/Dual%20ISH-IHC%20Pronin-SMA%20antibody%20and%20Olfr558%20RNAscope_0.jpg?itok=nt1KRE3G" width="765" height="451" alt="Dual ISH-IHC. Detection of alpha-smooth muscle actin (SMA) using IHC (green) and Olfr558 using RNAscope technology (red)"><p class="slider-below-text"> Dual ISH-IHC. Detection of alpha-smooth muscle actin (SMA) using IHC (green) and Olfr558 using RNAscope technology (red)</p></div></div><style type="text/css"> .loader-image-gallery { position: absolute; left: 0px; top: 0px; width: 100%; height: 100%; z-index: 9999; background: url('/sites/default/files/loading_apple.gif') 50% 35% no-repeat rgb(249,249,249); background-size: 100px; opacity: .8; } .sample-image { position: relative; } </style> <script type="text/javascript"> jQuery(document).ready(function($) { // $('#film_roll').hide(); function preloader(){ $('.loader-image-gallery').hide(); $('#film_roll').show(); } window.onload = preloader; }); </script> </div><div class="profile-content application_node_53409"><div class="field field-name-body field-type-text-with-summary field-label-hidden"><div class="field-items"><div class="field-item even" property="content:encoded"><h4><span style="font-size:18px;"><strong>NEW! <span class="ui-provider a b c d e f g h i j k l m n o p q r s t u v w x y z ab ac ae af ag ah ai aj ak" dir="ltr">Introducing our <a href="https://acdbio.com/PretreatmentOptions">protease-free pretreatment workflow</a> for sequential RNA-Protein detection</span></strong></span></h4><p> </p><p>Analysis of both RNA and protein provides invaluable information for understanding the regulation of expression. Both ISH and IHC technologies share common features that make them unique amongst the protein and RNA detection technologies.</p><p> - Morphological context: detection of RNA and protein while conserving the cell and tissue structural integrity</p><p> - Similar workflows from sample fixation, pre-treatment, hybridization, signal amplification and data analysis.</p><p>For labs that require high throughput and automated RNA <em>in situ</em> hybridization (ISH) analysis, ACD has partnered with <a href="https://acdbio.com/automated-assay-leica">Leica Biosystems</a> and <a href="https://acdbio.com/automated-assay-ventana">Roche Tissue Diagnostics</a>, Inc to deliver RNAscope™ ISH automated assays. Both automated stainers offering the possibility to combine <em>in situ</em> hybridization and immunohistochemistry on the same slide.</p><p>Due to the multiple benefits of running both ISH and IHC and the robustness of the RNAscope assay, some customers developed a dual ISH-IHC protocol to combine both assays on the same sample slide. We conducted an interview with 4 researchers who developed their own dual ISH-IHC method to provide you some insights into their protocol.</p><p>Download the interview and learn from these experts how to develop a dual ISH-IHC protocol.</p><div class="row"><div class="col-md-7"><p>Interview with:</p><ul><li><p><strong>Kristie Wetzel</strong>, Novartis Institute for</p><p>BioMedical Research, Cambridge, MA, USA.</p></li><li><p><strong>Dr Bradley Spencer-Dene</strong>, The Francis Crick</p><p>Institute, London, UK.</p></li><li><p><strong>Wanda Wang</strong>, University of Pittsburgh, PA,</p><p>USA.</p></li><li><p><strong>Mike Millar</strong>, Queen’s Medical Research</p><p>Institute, University of Edinburgh,</p><p>Edinburgh, UK.</p></li></ul></div><div class="col-md-5"><div class="download-pdf"><p><img alt="" src="/sites/default/files/dual-ish-ihc-interview.jpg" /></p><p><a class="down-pdf-btn" href="https://acdbio.com/system/files_force/gated/MK_51_024_Dual_ISH_IH_Innovation_in_the_spotlight..pdf?download=1">Download Interview</a></p></div></div></div></div></div></div><div class="view view-application view-id-application view-display-id-block_8 view-dom-id-c55d25c051fbaa2df238d5f77eb34ded"><div class="view-content"><div class="row immuno-watch"><h3 style="font-size: 24px;font-weight: 500;margin-left: 15px;margin-bottom: 0">Recorded Webinar</h3><div class="col-md-6"><p><em>Application of combined RNA in situ hybridization and multiplex immunohistochemistry to unravel tumor heterogeneity in prostate cancer.</em><br />Presented by Dr Nallasivam Palanisamy and Dr Courtney Anderson</p> <a class="immu-watch-link" href="https://www.labroots.com/ms/webinar/application-combined-rna-situ-hybridization-multiplex-immunohistochemistry-unravel-tumor-het"> Watch Webinar </a></div><div class="col-md-6"> <img src="https://acdbio.com/sites/default/files/dual-ihc.png" /></div></div></div></div><div class="view view-application view-id-application view-display-id-block_9 view-dom-id-c9c3a760deb09b5d3c0c5abbe156075d"><div class="view-content"><style type="text/css"> .view-id-application.view-display-id-block_9 .row.immuno-watch { margin-top: 15px; margin-bottom: 15px; } </style></div></div></div><div class="sample-image"><div class="profile-bottom"><h2>ISH-IHC App Note</h2><p>Learn more in our RNAscope ISH-IHC Application note.</p><div class="row review"><div class="col-md-6"> <img src="/sites/default/files/Flyer2.jpg"></div><div class="col-md-6"><h2>Fill out the form below to download the application note:</h2><div class="marketo-container download-pdf-app"><form id="mktoForm_9726" rel="9726"> </form> <a href="" class="down-pdf-btn marketo-popup">Download Pdf</a></div></div><div class="clear"></div></div></div></div><div class="spotlight"><div class="profile-bottom spot"><h2><span style="font-size:22px;">Related Publications</span></h2><p>Download our latest Dual ISH-IHC publication list with more than 300 publications.</p><p class="download-pdf"><a href="https://acdbio.com/system/files_force/gated/L_784.pdf?download=1">Download Publication list</a></p></div><div class="profile-bottom publication"><h2><span style="font-size:22px;">Related Protocol</span></h2><p>Download our RNAscope LS Multiplex Fluorescent Assay Combined with Immunofluorescence Technical Note.</p><p class="download-pdf"><a href="https://acdbio.com/system/files_force/gated/322818_TN_LS_Multiplex_and_immunofluorescence_02282017.pdf?download=1">Download Technical Note</a></p><p class="download-pdf">Download our RNAscope Multiplex Fluorescent Assay Combined with Immunofluorescence Technical Note</p><p class="download-pdf"><a href="https://acdbio.com/system/files_force/gated/TN_323100_TechNote_Dual_ISH_IHC_manual_Multiplex_Fluorescent_V2_06222017.pdf.pdf?download=1">Download Technical Note</a></p><p class="download-pdf">Download our RNAscope RED Assay and Immunofluorescence Technical Note</p><p class="download-pdf"><a href="https://acdbio.com/system/files_force/gated/TN_322350_TechNote_dual_ISH_IHC_manual_RNAscope_red__06192017.pdf.pdf?download=1">Download Technical Note</a></p><p class="download-pdf"> </p></div></div></div></div></div></div></div> <script type="text/javascript"> jQuery(document).ready(function($) { var windowsize = $(window).width(); 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type="button" id="closeInvite" class="embeddedServiceCloseIcon" aria-label="Exit invitation">×</button> </div> <div class="embeddedServiceInvitationBody"> <p id="snapins_bodytext">How can we help you?</p> </div> <div class="embeddedServiceInvitationFooter" aria-describedby="snapins_bodytext"> <button type="button" class="embeddedServiceActionButton" id="rejectInvite">Close</button> <button type="button" class="embeddedServiceActionButton" id="acceptInvite">Start Chat</button> </div> </div> <style type='text/css'> #snapins_invite { background-color: #FFFFFF; font-family: "Arial", sans-serif; overflow: visible; border-radius: 8px; visibility: hidden; } .embeddedServiceInvitation { background-color: transparent; max-width: 290px; max-height: 210px; -webkit-box-shadow: 0 7px 12px rgba(0,0,0,0.28); -moz-box-shadow: 0 7px 12px rgba(0,0,0,0.28); box-shadow: 0 7px 12px rgba(0,0,0,0.28); } @media only screen and (min-width: 48em) { /*mobile*/ .embeddedServiceInvitation { max-width: 332px; max-height: 210px; } } .embeddedServiceInvitation > .embeddedServiceInvitationHeader { width: inherit; height: 32px; line-height: 32px; padding: 10px; color: #FFFFFF; background-color: #222222; overflow: initial; display: flex; justify-content: space-between; align-items: stretch; border-top-left-radius: 8px; border-top-right-radius: 8px; } .embeddedServiceInvitationHeader #embeddedServiceAvatar { width: 32px; height: 32px; border-radius: 50%; } .embeddedServiceInvitationHeader .embeddedServiceTitleText { font-size: 18px; color: #FFFFFF; overflow: hidden; word-wrap: normal; white-space: nowrap; text-overflow: ellipsis; align-self: stretch; flex-grow: 1; max-width: 100%; margin: 0 12px; } .embeddedServiceInvitationHeader .embeddedServiceCloseIcon { border: none; border-radius: 3px; cursor: pointer; position: relative; bottom: 3%; background-color: transparent; width: 32px; height: 32px; font-size: 23px; color: #FFFFFF; } .embeddedServiceInvitationHeader .embeddedServiceCloseIcon:focus { outline: none; } .embeddedServiceInvitationHeader .embeddedServiceCloseIcon:focus::before { content: " "; position: absolute; top: 11%; left: 7%; width: 85%; height: 85%; background-color: rgba(255, 255, 255, 0.2); border-radius: 4px; pointer-events: none; } .embeddedServiceInvitationHeader .embeddedServiceCloseIcon:active, .embeddedServiceCloseIcon:hover { background-color: #FFFFFF; color: rgba(0,0,0,0.7); opacity: 0.7; } .embeddedServiceInvitation > .embeddedServiceInvitationBody { background-color: #FFFFFF; max-height: 110px; min-width: 260px; margin: 0 8px; font-size: 14px; line-height: 20px; overflow: auto; } .embeddedServiceInvitationBody p { color: #333333; padding: 8px; margin: 12px 0; } .embeddedServiceInvitation > .embeddedServiceInvitationFooter { width: inherit; color: #FFFFFF; text-align: right; background-color: #FFFFFF; padding: 10px; max-height: 50px; border-bottom-left-radius: 8px; border-bottom-right-radius: 8px; } .embeddedServiceInvitationFooter > .embeddedServiceActionButton { font-size: 14px; max-height: 40px; border: none; border-radius: 4px; padding: 10px; margin: 4px; text-align: center; text-decoration: none; display: inline-block; cursor: pointer; } .embeddedServiceInvitationFooter > #acceptInvite { background-color: #005290; color: #FFFFFF; } .embeddedServiceInvitationFooter > #rejectInvite { background-color: #FFFFFF; color: #005290; } </style>   <input type="hidden" name="liveagent.prechat.findorcreate.map.doFind:Contact" value="LastName,true;FirstName,true;Email,true" /> <input type="hidden" name="liveagent.prechat.findorcreate.map.isExactMatch:Contact" value="LastName,true;FirstName,true;Email,true" /> <input type="hidden" name="liveagent.prechat.findorcreate.map.doCreate:Contact" value="LastName,true;FirstName,true;Email,true" /> <script type='text/javascript'> (function() { document.getElementById('closeInvite').onclick = function() { embedded_svc.inviteAPI.inviteButton.rejectInvite(); }; document.getElementById('rejectInvite').onclick = function() { embedded_svc.inviteAPI.inviteButton.rejectInvite(); }; // use this API call to reject invitations document.getElementById('acceptInvite').onclick = function() { embedded_svc.liveAgentAPI.startChat({ directToButtonRouting: { buttonId: "5733t000000cVk9" }, extraPrechatInfo: [], extraPrechatFormDetails: [] }); }; // use this API call to start chat from invitations document.addEventListener('keyup', function(event) { if (event.keyCode == 27) { embedded_svc.inviteAPI.inviteButton.rejectInvite(); }}) })(); </script>  <script type='text/javascript' src='https://service.force.com/embeddedservice/5.0/esw.min.js'></script> <script type='text/javascript'> var initESW = function(gslbBaseURL) { embedded_svc.settings.displayHelpButton = true; //Or false embedded_svc.settings.language = ''; //For example, enter 'en' or 'en-US' embedded_svc.settings.defaultMinimizedText = 'Chat now'; //(Defaults to Chat with an Expert) embedded_svc.settings.disabledMinimizedText = 'Leave a message'; //(Defaults to Agent Offline) embedded_svc.settings.autoOpenPostChat = true; //embedded_svc.settings.loadingText = ''; //(Defaults to Loading) //embedded_svc.settings.storageDomain = 'yourdomain.com'; //(Sets the domain for your deployment so that visitors can navigate subdomains during a chat session) // Settings for Chat embedded_svc.settings.directToButtonRouting = function (prechatFormData) { if (prechatFormData[4].value === "Technical Service") return "5733t000000XZAM"; if (prechatFormData[4].value === "Customer Care") return "5733t000000XZAH"; }; embedded_svc.settings.prepopulatedPrechatFields = { OmniURL__c: window.location.href, ChooseaDepartment__c: 'Technical Service' }; //Sets the auto-population of pre-chat form fields embedded_svc.settings.extraPrechatFormDetails = [{ "label":"Source Domain", "value":window.location.href, "transcriptFields":[ "Source_Domain__c" ], "displayToAgent":true }]; //embedded_svc.settings.fallbackRouting = []; //An array of button IDs, user IDs, or userId_buttonId embedded_svc.settings.offlineSupportMinimizedText = 'Leave a message'; //(Defaults to Contact Us) embedded_svc.settings.enabledFeatures = ['LiveAgent']; embedded_svc.settings.entryFeature = 'LiveAgent'; embedded_svc.init( 'https://biotechne.my.salesforce.com', 'https://biotechne.my.salesforce-sites.com/LiveAgent', gslbBaseURL, '00D500000006gkl', 'ACD_Chat_Automated', { baseLiveAgentContentURL: 'https://c.la1-c1-ia5.salesforceliveagent.com/content', deploymentId: '572380000004CqL', buttonId: '5733t000000cVk9', baseLiveAgentURL: 'https://d.la1-c1-ia5.salesforceliveagent.com/chat', eswLiveAgentDevName: 'EmbeddedServiceLiveAgent_Parent04I3t000000Y7zbEAC_175bb741b99', isOfflineSupportEnabled: true } ); }; if (!window.embedded_svc) { var s = document.createElement('script'); s.setAttribute('src', 'https://biotechne.my.salesforce.com/embeddedservice/5.0/esw.min.js'); s.onload = function() { initESW(null); }; document.body.appendChild(s); } else { initESW('https://service.force.com'); } </script> <script src="//rum-static.pingdom.net/pa-5c133f6e1872b50016000024.js" async></script> <script type="text/javascript">window.NREUM||(NREUM={});NREUM.info={"beacon":"bam.nr-data.net","licenseKey":"NRJS-dd6144c4de227d09eb4","applicationID":"1022125825","transactionName":"YFxbZkpUXBYEVkYNWVkWbEBRGlsLAVBKSkZfSQ==","queueTime":0,"applicationTime":416,"atts":"TBtYEAJOTxg=","errorBeacon":"bam.nr-data.net","agent":""}</script></body> </html>