CRISPR-Cas9 Antibody Pack (NBP2-52986) by Novus, Part of Bio-Techne

<!DOCTYPE html> <html lang="en" dir="ltr" prefix="og: https://ogp.me/ns#"> <head> <meta charset="utf-8" /> <noscript><style>form.antibot * :not(.antibot-message) { display: none !important; }</style> </noscript><meta name="applicable-device" content="pc,mobile" /> <link rel="canonical" href="https://www.bio-techne.com/p/antibodies/crispr-cas9-antibody-pack_nbp2-52986" /> <meta property="og:site_name" content="Bio-Techne" /> <meta property="og:type" content="Website" /> <meta property="og:url" content="https://www.bio-techne.com/" /> <meta name="Generator" content="Drupal 10 (https://www.drupal.org)" /> <meta name="MobileOptimized" content="width" /> <meta name="HandheldFriendly" content="true" /> <meta name="viewport" content="width=device-width, initial-scale=1.0" /> <script type="application/ld+json">{ "@context": "https://schema.org", "@graph": [ { "@type": "WebPage", "@id": "https://www.bio-techne.com/p/antibodies/crispr-cas9-antibody-pack_nbp2-52986", "breadcrumb": { "@type": "BreadcrumbList", "itemListElement": [ { "@type": "ListItem", "position": 1, "name": "Home", "item": "https://www.bio-techne.com/" }, { "@type": "ListItem", "position": 2, "name": "CRISPR-Cas9", "item": "https://www.bio-techne.com/t/crispr-cas9" }, { "@type": "ListItem", "position": 3, "name": "CRISPR-Cas9 Antibodies", "item": "https://www.bio-techne.com/t/crispr-cas9/antibodies" } ] } } ] }</script> <meta http-equiv="x-dns-prefetch-control" content="on" /> <meta http-equiv="x-ua-compatible" content="ie=edge" /> <meta name="keywords" content="" /> <meta name="description" content="Purchase Novus Biologicals CRISPR-Cas9 Antibody Pack. Backed by our 100% Guarantee." /> <script type="application/ld+json" id="json-ld">[{"@context":"http://schema.org","@type":"Product","additionalProperty":[],"aggregateRating":[],"alternateName":"","brand":{"@type":"Brand","name":"Novus Biologicals"},"description":"","image":[{"@type":"ImageObject","url":"https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NBP2-52986-img0004.jpg","description":"Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986]"},{"@type":"ImageObject","url":"https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0005.jpg","description":"Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]"},{"@type":"ImageObject","url":"https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Chromatin-Immunoprecipitation-ChIP-NBP2-52986-img0001.jpg","description":"Chromatin Immunoprecipitation (ChIP): CRISPR-Cas9 Antibody Pack [NBP2-52986] - ChIP analysis of CRISPR-Cas9 antibody clones 7A9-3A3 and 6G12 on NIH3T3 cells which were stably expressing GFP-H2B, nuclease dead Cas9 (Flag tagged), and a GFP-targeting gRNA. The cells were fixed with formaldehyde, harvested and sonicated to get 200-500bp DNA fragments. 50ug chromatin was incubated over night at 4C with the indicated antibodies (200ul hybridoma supernatants of clones 7A9 and 6G12; 5ug Flag antibody) followed by incubation with protein G beads for 3h at 4C. After washing chromatin was eluted from the beads and crosslinking was reversed over night at 65C. After a proteinase K digestion step, DNA was separated using phenol/chloroform/isoamyl alcohol, precipitated with ethanol/sodium acetate and dissolved in water. For qPCR, primers either targeting the GFP gene or as negative control non-targeted regions (Ppap2c +7122 and Prkcd +24069 from transcription start) were used."},{"@type":"ImageObject","url":"https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0003.jpg","description":"Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]"},{"@type":"ImageObject","url":"https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunoprecipitation-NBP2-52986-img0002.jpg","description":"Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986]"}],"isRelatedTo":[],"itemcondition":"new","mpn":"NBP2-52986","name":"CRISPR-Cas9 Antibody Pack","productID":"NBP2-52986","review":[],"sku":"NBP2-52986","url":"https://www.bio-techne.com/p/antibodies/crispr-cas9-antibody-pack_nbp2-52986"},{"@context":"http://schema.org","@type":"AskAction","question":[]}]</script> <link rel="alternate" hreflang="en" 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class="col sub-menu-wrapper"> <div class="sub-menu-header"> Product & Instrument Tools<span class="icon-keyboard_arrow_down submenu-caret"></span> <hr/> <div class="menu-children-wrap sub-menu-box"> <a href="/resources/cofa-finder-tool" class="dropdown-item text-wrap" data-drupal-link-system-path="resources/cofa-finder-tool">Certificate of Analysis (COA) Finder Tool</a> <a href="/resources/calculators" class="dropdown-item text-wrap" data-drupal-link-system-path="node/7416">Calculators</a> <a href="/resources/flow-cytometry-panel-builder" class="dropdown-item text-wrap" data-drupal-link-system-path="node/8821">Flow Cytometry Panel Builder</a> <a href="/luminex-assay-customization-tool" class="dropdown-item text-wrap" data-drupal-link-system-path="luminex-assay-customization-tool">Luminex Assay Customization Tool</a> <a href="/resources/product-insert-lookup-tool" class="dropdown-item text-wrap" data-drupal-link-system-path="resources/product-insert-lookup-tool">Product Insert Lookup 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data-time_datapoints="-0.25974488258362" data-time_template="0.25974488258362" data-time_post_epcc="0.97506403923035" data-time_epcc="-0.25974488258362" data-render_time="Friday 4th of October 2024 06:34:08 PM" data-fallback="FALSE" data-time_to_process_data="0.090008020401001" class="px-4 container" class="container"> <div id="above-fold-section"> <div class="mb-2 o__product-summary"> <div id="header-subheader" data-section="section-base--header-subheader"> <div class="validation-badges-header-wrapper"> <a class="biological-validation mr-3 d-inline-flex mt-1" href="/reagents/antibodies/antibody-validation" target="_blank" title="Biological Validation"></a> </div> <div class="row" id="pp-page-title-brand-wrapper"> <div class="col-lg-9 col-md-12"> <h1 class="font-weight-bold"> CRISPR-Cas9 Antibody Pack </h1> <h2 class="font-weight-normal h4"> Novus Biologicals, part of Bio-Techne <span id="product-code"> | Catalog # <span class="font-weight-bold">NBP2-52986</span> </span> </h2> <div 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</div> <div class="row border-top mb-2" id="images-orderdetail-wrapper"> <div class="col-xl-6 mt-2" id="images-wrapper" data-section="image_summary"> <div id="image-tree" data-section="component-product-image-tree" class="vertical lightbox" data-mdb-zoom-effect="true"> <div id="product-image-gallery" class="row"> <div class="col-md-2 d-none d-md-block"> <ul class="list-inline m-0"> <li data-id="1" class="product-image-lightbox border rounded mb-2 list-inline-item"> <div class="text-center mt-1"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0005.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0005.jpg" alt="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986] - Hela cells and Hela cells expressing Flag-tagged S.pyogenes Cas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/ul Doxycyclin and lysed under native conditions. 30ug of whole cell lysate per lane was separated by 7.5% SDS-PAGE, transfered to nitrocellulose membrane and incubated with 6G12 hybridoma supernatant (diluted 1:100) at 4C o/n. After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films. Prestained marker bands were visualized with Blue Marker Antibody (NBP2-33376). A 1 min exposure is shown. Note the specific band of CRISPR-Cas9 with a molecular weight of 158.4 kDa." data-badges='' data-mdb-img="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0005.jpg" data-id="0" data-application="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0005.jpg" class="lazy" /> </div> </li> <li data-id="2" class="product-image-lightbox border rounded mb-2 list-inline-item"> <div class="text-center mt-1"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Chromatin-Immunoprecipitation-ChIP-NBP2-52986-img0001.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Chromatin-Immunoprecipitation-ChIP-NBP2-52986-img0001.jpg" alt="" title="" data-caption="Chromatin Immunoprecipitation (ChIP): CRISPR-Cas9 Antibody Pack [NBP2-52986] - ChIP analysis of CRISPR-Cas9 antibody clones 7A9-3A3 and 6G12 on NIH3T3 cells which were stably expressing GFP-H2B, nuclease dead Cas9 (Flag tagged), and a GFP-targeting gRNA. The cells were fixed with formaldehyde, harvested and sonicated to get 200-500bp DNA fragments. 50ug chromatin was incubated over night at 4C with the indicated antibodies (200ul hybridoma supernatants of clones 7A9 and 6G12; 5ug Flag antibody) followed by incubation with protein G beads for 3h at 4C. After washing chromatin was eluted from the beads and crosslinking was reversed over night at 65C. After a proteinase K digestion step, DNA was separated using phenol/chloroform/isoamyl alcohol, precipitated with ethanol/sodium acetate and dissolved in water. For qPCR, primers either targeting the GFP gene or as negative control non-targeted regions (Ppap2c +7122 and Prkcd +24069 from transcription start) were used." data-badges='' data-mdb-img="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Chromatin-Immunoprecipitation-ChIP-NBP2-52986-img0001.jpg" data-id="1" data-application="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Chromatin-Immunoprecipitation-ChIP-NBP2-52986-img0001.jpg" class="lazy" /> </div> </li> <li data-id="3" class="product-image-lightbox border rounded mb-2 list-inline-item"> <div class="text-center mt-1"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0003.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0003.jpg" alt="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986] - WB analysis of Cas9 protein in lysate of CRISPR-Cas9 transfected 293T cells using anti-Cas9 antibody clone 7A9-3A3 at 2ug/ml concentration." data-badges='' data-mdb-img="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0003.jpg" data-id="2" data-application="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0003.jpg" class="lazy" /> </div> </li> <li data-id="4" class="product-image-lightbox border rounded mb-2 list-inline-item"> <div class="text-center mt-1"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunoprecipitation-NBP2-52986-img0002.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunoprecipitation-NBP2-52986-img0002.jpg" alt="Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986] - HEK293T expressing N-terminally Flag-tagged S.pyogenes Cas9 were lysed 72h post transfection by resuspending the cells in Hunt buffer and subjecting to 3 freeze-thaw cycles in liquid nitrogen/ice. Proteins were immunoprecipitated from 100ug of whole cell lysate for 1h at 4C with Cas9 supernatant followed by incubation for 1h at 4C with a 1:1 mixture of protein A/G sepharose beads, or for 2h at 4C with Cas9 ab crosslinked to a 1:1 mixture of protein A/G sepharose beads. Beads were washed 2x with Hunt buffer and 1x with TBS. Bound proteins were eluted by boiling in Laemmli, separated by SDS-PAGE and transferred to nitrocellulose. Membrane was blocked, incubated with Cas9 ab, incubated with HRP anti-mouse secondary. *IgG heavy chain." data-badges='' data-mdb-img="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunoprecipitation-NBP2-52986-img0002.jpg" data-id="3" data-application="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunoprecipitation-NBP2-52986-img0002.jpg" class="lazy" /> </div> </li> </ul> </div> <div class="col-md-10 border rounded d-flex align-items-center justify-content-center product-image-lightbox"> <div class="text-center my-2" data-brand="novus"> <img src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NBP2-52986-img0004.jpg" alt="Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986] - Hela cells or Hela cells expressing Flag-tagged SpCas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/ul Doxycyclin, fixed and permeabilized with Methanol/Acetone and blocked in 2% BSA in PBS for 2 hours at RT. Cells were stained with 6G12 hybridoma supernatant (diluted 1:10) at 4C o/n, followed by incubation with anti mouse-AF488 coupled secondary antibody for 1 h at RT. Nuclei were counter-stained with Hoechst 33342." data-badges='' data-id="0" width="250" height="250" fetchpriority="high" as="image" class="search_result_thumbnail_image active" /> </div> </div> </div> </div> </div> <div class="col-xl-6 mt-2" id="orderdetails-wrapper" data-section="order_detail_block"> <div class="atc-container"> <div class="atc-block" data-type="ds" data-blockid="ProductCart" > <form action="/commerce/addtocart" method="post" class="add-to-cart-block" data-productcode="NBP2-52986"> <input type="hidden" name="print-quote" class="print-quote-input" value=""> <div class="commerce-atc-full-header atc-header col-four row border-bottom pb-1 m-0"> <div class="commerce-atc-catnum atc-head col-4 col-md-3 font-weight-bold p-0">Catalog #</div> <div class="commerce-atc-availability atc-head col-md-4 p-0 font-weight-bold d-none d-md-block">Availability</div> <div class="commerce-atc-price atc-head col-4 col-md-3 p-0 font-weight-bold">Size / Price</div> <div 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d-inline jump-anchor " href="#specific-notices"> Specific Notices</a> </li> </ul> </div> <div id="key_details" class="py-5 bt-pp-border-bottom" data-section="section-base--key-details"> <h3>Key Product Details</h3> <div class="mt-4 py-2 container"> <div class="row product-data"> <div class="col-lg-3 font-weight-bold product-data-label py-2 align-top text-bt-dark-blue"> <h4>Species</h4> </div> <div class="col-lg-9 product-data-value py-2 align-top text-break"> Bacteria </div> </div> <div class="row product-data"> <div class="col-lg-3 font-weight-bold product-data-label py-2 align-top text-bt-dark-blue"> <h4>Applications</h4> </div> <div class="col-lg-9 product-data-value py-2 align-top text-break"> Chromatin Immunoprecipitation, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Simple Western, Western Blot </div> </div> </div> </div> <div class="border-top border-bottom recommended-products-section" id="recommendation-viewed" data-product_code="NBP2-52986" data-type=viewed></div> <div id="product-summary" class="py-5 bt-pp-border-bottom" data-section="section-generic--product-summary"> <div> <h3>Product Summary for CRISPR-Cas9 Antibody Pack </h3> </div> <div class="mt-2 py-2"> <div class="row product-data"> <div class="col-lg-12 product-data-value py-2 align-top text-break"> This pack contains 1 vial each of: NBP2-36440SS (0.025 mL), NBP2-52398SS (0.025 mL). </div> </div> </div> </div> <div id="product-specifications" class="bt-pp-border-bottom py-5" data-section="section-base--product-specifications"> <h3>Product Specifications</h3> <div class="mt-4 py-2 container"> <div class="row product-data"> <div class="col-lg-3 font-weight-bold product-data-label py-2 align-top text-bt-dark-blue"> <h4>Application Notes</h4> </div> <div class="col-lg-9 product-data-value py-2 align-top text-break"> See individual datasheets of components for their validated applications </div> </div> <div class="row product-data"> <div class="col-lg-3 font-weight-bold product-data-label py-2 align-top text-bt-dark-blue"> <h4>Reactivity Notes</h4> </div> <div class="col-lg-9 product-data-value py-2 align-top text-break"> See individual datasheets of components for their validated species </div> </div> </div> </div> <div id="scientific-data" class="bt-pp-border-bottom py-5" data-anchor-label="Scientific Data" data-section="section-base--scientific-data"> <div> <h3>Scientific Data Images for CRISPR-Cas9 Antibody Pack </h3> </div> <div class="product-data-wrapper mt-5" id="scientific-data-table" data-scientific-count="2"> <div> <div class="row m___product-data-examples my-4" data-section="product-image-divs"> <div class="col-md-2 border-0 mb-3"> <div class="image-slider-container"> <div class="product-image-lightbox" data-brand="novus"> <div class="text-center border rounded p-2 item d-flex align-items-center justify-content-center bg-white active"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NBP2-52986-img0004.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunocytochemistry-Immunofluorescence-NBP2-52986-img0004.jpg" alt="Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986] - Hela cells or Hela cells expressing Flag-tagged SpCas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/ul Doxycyclin, fixed and permeabilized with Methanol/Acetone and blocked in 2% BSA in PBS for 2 hours at RT. Cells were stained with 6G12 hybridoma supernatant (diluted 1:10) at 4C o/n, followed by incubation with anti mouse-AF488 coupled secondary antibody for 1 h at RT. Nuclei were counter-stained with Hoechst 33342." data-badges="" data-application="Immunocytochemistry/ Immunofluorescence" width="250" height="250" class="search_result_thumbnail_image lazy" /> <div class="slider-open"> <div class="count-container text-center px-2 py-2 border-top border-left rounded-left"> <span class="icon-icon-image-camera-plus align-middle"></span> </div> </div> </div> </div> </div> </div> <div class="col-md-9 border-0 align-top"> <h4 class="font-weight-bold">Immunocytochemistry/ Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986]</h4> Immunocytochemistry/Immunofluorescence: CRISPR-Cas9 Antibody Pack [NBP2-52986] - Hela cells or Hela cells expressing Flag-tagged SpCas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/ul Doxycyclin, fixed and permeabilized with Methanol/Acetone and blocked in 2% BSA in PBS for 2 hours at RT. Cells were stained with 6G12 hybridoma supernatant (diluted 1:10) at 4C o/n, followed by incubation with anti mouse-AF488 coupled secondary antibody for 1 h at RT. Nuclei were counter-stained with Hoechst 33342. </div> </div> </div> <div> <div class="row m___product-data-examples my-4" data-section="product-image-divs"> <div class="col-md-2 border-0 mb-3"> <div class="image-slider-container"> <div class="product-image-lightbox" data-brand="novus"> <div class="text-center border rounded p-2 item d-flex align-items-center justify-content-center bg-white active"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0005.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0005.jpg" alt="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986] - Hela cells and Hela cells expressing Flag-tagged S.pyogenes Cas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/ul Doxycyclin and lysed under native conditions. 30ug of whole cell lysate per lane was separated by 7.5% SDS-PAGE, transfered to nitrocellulose membrane and incubated with 6G12 hybridoma supernatant (diluted 1:100) at 4C o/n. After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films. Prestained marker bands were visualized with Blue Marker Antibody (NBP2-33376). A 1 min exposure is shown. Note the specific band of CRISPR-Cas9 with a molecular weight of 158.4 kDa." data-badges="" data-application="Western Blot" width="250" height="250" class="search_result_thumbnail_image lazy" /> <div class="slider-open"> <div class="count-container text-center px-2 py-2 border-top border-left rounded-left"> <span class="icon-icon-image-camera-plus align-middle"></span> </div> </div> </div> </div> </div> </div> <div class="col-md-9 border-0 align-top"> <h4 class="font-weight-bold">Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]</h4> Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986] - Hela cells and Hela cells expressing Flag-tagged S.pyogenes Cas9 under the control of the PTight (Tet-ON) promoter were treated for 24h with 1ug/ul Doxycyclin and lysed under native conditions. 30ug of whole cell lysate per lane was separated by 7.5% SDS-PAGE, transfered to nitrocellulose membrane and incubated with 6G12 hybridoma supernatant (diluted 1:100) at 4C o/n. After washing, the membranes were incubated with secondary HRP-coupled antibody and bands were visualized by ECL and exposure of X-ray films. Prestained marker bands were visualized with Blue Marker Antibody (NBP2-33376). A 1 min exposure is shown. Note the specific band of CRISPR-Cas9 with a molecular weight of 158.4 kDa. </div> </div> </div> <div> <div class="row m___product-data-examples my-4" data-section="product-image-divs"> <div class="col-md-2 border-0 mb-3"> <div class="image-slider-container"> <div class="product-image-lightbox" data-brand="novus"> <div class="text-center border rounded p-2 item d-flex align-items-center justify-content-center bg-white active"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Chromatin-Immunoprecipitation-ChIP-NBP2-52986-img0001.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Chromatin-Immunoprecipitation-ChIP-NBP2-52986-img0001.jpg" alt="" title="" data-caption="Chromatin Immunoprecipitation (ChIP): CRISPR-Cas9 Antibody Pack [NBP2-52986] - ChIP analysis of CRISPR-Cas9 antibody clones 7A9-3A3 and 6G12 on NIH3T3 cells which were stably expressing GFP-H2B, nuclease dead Cas9 (Flag tagged), and a GFP-targeting gRNA. The cells were fixed with formaldehyde, harvested and sonicated to get 200-500bp DNA fragments. 50ug chromatin was incubated over night at 4C with the indicated antibodies (200ul hybridoma supernatants of clones 7A9 and 6G12; 5ug Flag antibody) followed by incubation with protein G beads for 3h at 4C. After washing chromatin was eluted from the beads and crosslinking was reversed over night at 65C. After a proteinase K digestion step, DNA was separated using phenol/chloroform/isoamyl alcohol, precipitated with ethanol/sodium acetate and dissolved in water. For qPCR, primers either targeting the GFP gene or as negative control non-targeted regions (Ppap2c +7122 and Prkcd +24069 from transcription start) were used." data-badges="" data-application="Chromatin Immunoprecipitation (ChIP)" width="250" height="250" class="search_result_thumbnail_image lazy" /> <div class="slider-open"> <div class="count-container text-center px-2 py-2 border-top border-left rounded-left"> <span class="icon-icon-image-camera-plus align-middle"></span> </div> </div> </div> </div> </div> </div> <div class="col-md-9 border-0 align-top"> Chromatin Immunoprecipitation (ChIP): CRISPR-Cas9 Antibody Pack [NBP2-52986] - ChIP analysis of CRISPR-Cas9 antibody clones 7A9-3A3 and 6G12 on NIH3T3 cells which were stably expressing GFP-H2B, nuclease dead Cas9 (Flag tagged), and a GFP-targeting gRNA. The cells were fixed with formaldehyde, harvested and sonicated to get 200-500bp DNA fragments. 50ug chromatin was incubated over night at 4C with the indicated antibodies (200ul hybridoma supernatants of clones 7A9 and 6G12; 5ug Flag antibody) followed by incubation with protein G beads for 3h at 4C. After washing chromatin was eluted from the beads and crosslinking was reversed over night at 65C. After a proteinase K digestion step, DNA was separated using phenol/chloroform/isoamyl alcohol, precipitated with ethanol/sodium acetate and dissolved in water. For qPCR, primers either targeting the GFP gene or as negative control non-targeted regions (Ppap2c +7122 and Prkcd +24069 from transcription start) were used. </div> </div> </div> <span class="product-data-wrapper table" id="more-scientific-data" style="display:none"> <div class="row m___product-data-examples my-4" data-section="product-image-divs"> <div class="col-md-2 border-0 mb-3"> <div class="image-slider-container"> <div class="product-image-lightbox" data-brand="novus"> <div class="text-center border rounded p-2 item d-flex align-items-center justify-content-center bg-white active"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0003.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Western-Blot-NBP2-52986-img0003.jpg" alt="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986] - WB analysis of Cas9 protein in lysate of CRISPR-Cas9 transfected 293T cells using anti-Cas9 antibody clone 7A9-3A3 at 2ug/ml concentration." data-badges="" data-application="Western Blot" width="250" height="250" class="search_result_thumbnail_image lazy" /> <div class="slider-open"> <div class="count-container text-center px-2 py-2 border-top border-left rounded-left"> <span class="icon-icon-image-camera-plus align-middle"></span> </div> </div> </div> </div> </div> </div> <div class="col-md-9 border-0 align-top"> <h4 class="font-weight-bold">Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986]</h4> Western Blot: CRISPR-Cas9 Antibody Pack [NBP2-52986] - WB analysis of Cas9 protein in lysate of CRISPR-Cas9 transfected 293T cells using anti-Cas9 antibody clone 7A9-3A3 at 2ug/ml concentration. </div> </div> <div class="row m___product-data-examples my-4" data-section="product-image-divs"> <div class="col-md-2 border-0 mb-3"> <div class="image-slider-container"> <div class="product-image-lightbox" data-brand="novus"> <div class="text-center border rounded p-2 item d-flex align-items-center justify-content-center bg-white active"> <img src="data:image/png;base64,R0lGODlhAQABAAD/ACwAAAAAAQABAAACADs=" data-src="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunoprecipitation-NBP2-52986-img0002.jpg" data-srcset="https://resources.bio-techne.com/images/products/CRISPR-Cas9-Antibody-Pack-Immunoprecipitation-NBP2-52986-img0002.jpg" alt="Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986]" title="Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986]" data-caption="Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986] - HEK293T expressing N-terminally Flag-tagged S.pyogenes Cas9 were lysed 72h post transfection by resuspending the cells in Hunt buffer and subjecting to 3 freeze-thaw cycles in liquid nitrogen/ice. Proteins were immunoprecipitated from 100ug of whole cell lysate for 1h at 4C with Cas9 supernatant followed by incubation for 1h at 4C with a 1:1 mixture of protein A/G sepharose beads, or for 2h at 4C with Cas9 ab crosslinked to a 1:1 mixture of protein A/G sepharose beads. Beads were washed 2x with Hunt buffer and 1x with TBS. Bound proteins were eluted by boiling in Laemmli, separated by SDS-PAGE and transferred to nitrocellulose. Membrane was blocked, incubated with Cas9 ab, incubated with HRP anti-mouse secondary. *IgG heavy chain." data-badges="" data-application="Immunoprecipitation" width="250" height="250" class="search_result_thumbnail_image lazy" /> <div class="slider-open"> <div class="count-container text-center px-2 py-2 border-top border-left rounded-left"> <span class="icon-icon-image-camera-plus align-middle"></span> </div> </div> </div> </div> </div> </div> <div class="col-md-9 border-0 align-top"> <h4 class="font-weight-bold">Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986]</h4> Immunoprecipitation: CRISPR-Cas9 Antibody Pack [NBP2-52986] - HEK293T expressing N-terminally Flag-tagged S.pyogenes Cas9 were lysed 72h post transfection by resuspending the cells in Hunt buffer and subjecting to 3 freeze-thaw cycles in liquid nitrogen/ice. Proteins were immunoprecipitated from 100ug of whole cell lysate for 1h at 4C with Cas9 supernatant followed by incubation for 1h at 4C with a 1:1 mixture of protein A/G sepharose beads, or for 2h at 4C with Cas9 ab crosslinked to a 1:1 mixture of protein A/G sepharose beads. Beads were washed 2x with Hunt buffer and 1x with TBS. Bound proteins were eluted by boiling in Laemmli, separated by SDS-PAGE and transferred to nitrocellulose. Membrane was blocked, incubated with Cas9 ab, incubated with HRP anti-mouse secondary. *IgG heavy chain. </div> </div> </span> <div class="row"> <div class="col-md-2"> <a id="scientific-view-more" class="more"> View 2 more images </a> </div> </div> </div> </div> <div id="kit-contents" class="py-5 bt-pp-border-bottom" data-anchor-label="Kit Contents" data-section="section-base--kit-contents"> <h3>Kit Contents for CRISPR-Cas9 Antibody Pack </h3> <div class="product-data-value"> <ul><li><a href="/p/antibodies/crispr-cas9-antibody-6g12_nbp2-52398">NBP2-52398: CRISPR-Cas9 Antibody (6G12) - C-terminus - BSA Free</a></li><li><a href="/p/antibodies/crispr-cas9-antibody-7a9-3a3_nbp2-36440">NBP2-36440: CRISPR-Cas9 Antibody (7A9-3A3) - N-Terminus - BSA Free</a></li><li><a href="/p/secondary-antibodies/mouse-igg-horseradish-peroxidase-conjugated-antibody_haf007">HAF007: Mouse IgG Horseradish Peroxidase-conjugated Antibody</a></li></ul> </div> </div> <div id="preparation--storage" class="bt-pp-border-bottom py-5" data-section="section-base--preparation-storage"> <h3>Formulation, Preparation, and Storage</h3> <div class="mt-4 py-2 container"> <div class="row product-data"> <div class="col-lg-3 font-weight-bold product-data-label py-2 align-top text-bt-dark-blue"> <h4>Concentration</h4> </div> <div class="col-lg-9 product-data-value py-2 align-top text-break"> Concentration of individual antibodies may be found on the vial label. If unlisted please contact technical services. </div> </div> <div class="row product-data"> <div class="col-lg-3 font-weight-bold product-data-label py-2 align-top text-bt-dark-blue"> <h4>Shipping</h4> </div> <div class="col-lg-9 product-data-value py-2 align-top text-break"> The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. </div> </div> <div class="row product-data"> <div class="col-lg-3 font-weight-bold product-data-label py-2 align-top text-bt-dark-blue"> <h4>Storage</h4> </div> <div class="col-lg-9 product-data-value py-2 align-top text-break"> Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. </div> </div> </div> </div> <div id="background" class="py-5 bt-pp-border-bottom" data-section="section-base--background-info"> <h3>Background: CRISPR-Cas9</h3> <article class="product-data-wrapper"> Clustered regularly interspaced short palindromic repeats (CRISPRs) are derived from DNA fragments of bacteriophages that infect prokaryotes. When infected, the bacteria capture snips of DNA from the invading virus to create CRISPR arrays. During subsequent infections, the bacteria produce RNA segments from the CRISPR arrays to target the virus' DNA. CRISPR-associated protein 9 (Cas9) is RNA-guided, binds DNA, and is a cleaving enzyme that functions as an integral component of the bacterial CRISPR adaptive immune system that targets the virus' DNA to disable it (1). To check for sites complementary to the 20 base pair spacer region of the guide RNA (gRNA) of the CRISPR, Cas9 unwinds foreign DNA that invades the bacteria. If the DNA substrate is complementary to the gRNA, Cas9 cleaves the invading DNA, rendering the virus disabled. The presence of a 5'-NGG-3' protospacer adjacent motif (PAM) sequence immediately downstream of the target DNA (protospacer) is required for Cas9 cleavage of foreign DNA. As PAM is absent in bacterial CRISPR loci, cleavage of the host genome is avoided and provides a novel sequence for identification of foreign DNA by Cas9.<br/><br/>Using CRISPR-Cas9 technology, double-stranded DNA breaks may be induced within specific targeted genome sequences (target DNA; protospacer) for insertion or removal of DNA sequences for gene editing applications. To target a specific loci, a gRNA that will bind to a specific target sequence of DNA within a genome is created. The gRNA will recognize the DNA sequence, and the Cas9 enzyme will cleave the DNA at the targeted location. Once the targeted DNA is removed by Cas9, the cell's own DNA repair mechanism is used to insert or remove a DNA sequence for genomic editing. <br/><br/>Cas9 detection is used to confirm and evaluate CRISPR Cas9 gRNA transfection efficiency. Western blot analysis of CRISPR-Cas9 gRNA transfected cell lysates with Cas9 antibodies identifies the protein having a theoretical molecular weight of 160kDa. Broad areas of research are benefiting from CRISPR-Cas9 based gene editing tools including studies of basic immunity functions, genetic screening and disease treatment (2). Ethical concerns have led to many countries making it illegal to manipulate human germline cells or perform embryo genome editing. <br/><br/>References<br/><br/>1. Oakes, B. L., Fellmann, C., Rishi, H., Taylor, K. L., Ren, S. M., Nadler, D. C., . . . Savage, D. F. (2019). CRISPR-Cas9 Circular Permutants as Programmable Scaffolds for Genome Modification. Cell, 176(1-2), 254-267.e216. doi:10.1016/j.cell.2018.11.052<br/><br/>2. Chiou, S. H., Winters, I. P., Wang, J., Naranjo, S., Dudgeon, C., Tamburini, F. B., . . . Winslow, M. M. (2015). Pancreatic cancer modeling using retrograde viral vector delivery and in vivo CRISPR/Cas9-mediated somatic genome editing. Genes Dev, 29(14), 1576-1585. doi:10.1101/gad.264861.115 </article> <div id="background_wrapper" class="px-3 mt-2"> <div class="row product-data"> <div class=" col-md-3 product-data-label font-weight-bold p-3 align-top border"> <h4>Long Name</h4> </div> <div class=" col-md-9 product-data-value p-3 px-4 text-break border"> CRISPR-associated Protein 9 </div> </div> <div class="row product-data"> <div class=" col-md-3 product-data-label font-weight-bold p-3 align-top border"> <h4>Alternate Names</h4> </div> <div class=" col-md-9 product-data-value p-3 px-4 text-break border"> Cas9, CRISPR-associated endonuclease Cas9/Csn1, CRISPR-Cas9/Csn1, CRISPR/Cas9, csn1, SPy_1046, SPy1046, SpyCas9, CRISPR-associated protein 9 nuclease </div> </div> </div> <h4 class="mt-3">Additional CRISPR-Cas9 Products</h4> <ul class="list-unstyled"> <li><a href="/t/crispr-cas9" data-product="CRISPR-Cas9">All Products for CRISPR-Cas9 </a></li> <li><a href="/t/crispr-cas9/antibodies" data-application="CRISPR-Cas9 Primary Antibodies">CRISPR-Cas9 Primary Antibodies </a></li> <li><a href="/t/crispr-cas9/proteins-enzymes" data-application="CRISPR-Cas9 Proteins and Enzymes">CRISPR-Cas9 Proteins and Enzymes </a></li> </ul> </div> <div id="product-documents" data-section="section-base--product-documents"> <div id="accordion" class="border mb-4"> <div class="rounded-top bg-white"> <h3 class="m-0 p-3"> Product Documents for CRISPR-Cas9 Antibody Pack </h3> </div> <div class="vtab-section" data-section="Product Datasheets"> <div class="card-header rounded-top"> <a class="" data-toggle="collapse" data-target="#collapse_two" aria-expanded="true" aria-controls="collapse_two" id="product-datasheets-link"> <h5 class="mb-0"> Product Datasheets </h5> </a> </div> <div id="collapse_two" class="collapse show prod_datasheet_link" aria-labelledby="heading_two" data-parent="#accordion"> <div class="card-body"> <div class="row"> <div class="col-md-6"> <a href="https://resources.bio-techne.com/products/documents/datasheets/NBP2-52986.pdf" class="datasheet_link" target="_blank" rel="nofollow">Download Product Datasheets</a> </div> <div class="col-md-6 datasheet-pdf-result"> </div> </div> </div> </div> </div> <div class="card-header"> <a data-toggle="collapse" data-target="#collapse_one" aria-expanded="false" class="collapsed" aria-controls="collapse_one"> <h5 class="mb-0"> COA </h5> </a> </div> <div id="collapse_one" class="collapse " aria-labelledby="heading_one" data-parent="#accordion"> <div class="card-body"> <form novalidate="novalidate" data-drupal-selector="product-coa-form" action="#coa-form" method="post" id="product-coa-form" accept-charset="UTF-8"> <div class="row"> <div class="col-12"> <h2>Certificate of Analysis</h2> <p>To download a Certificate of Analysis, please enter a lot number in the search box below.</p> </div> </div> <section class="row"> <div class="col-md-6"> <form novalidate="novalidate" data-drupal-selector="product-coa-form"> <div class="form-row align-items-center"> <div class="col-md-10 my-1"> <input autocomplete="off" data-drupal-selector="form-3-bcuo2fcaa-idryq10p61pknakcqrwsiehakczhi9q" type="hidden" name="form_build_id" value="form-3-bcuO2fcAa-idRyQ10p61pkNaKcqRWsIEHakCzHi9Q" /> <input data-drupal-selector="edit-product-coa-form" type="hidden" name="form_id" value="product_coa_form" /> <input data-drupal-selector="edit-product-code" type="hidden" name="product_code" value="NBP2-52986" /> <div class="js-form-item form-item pb-3 js-form-type-textfield form-item-lot-number js-form-item-lot-number form-no-label"> <input autocomplete="off" class="form-control cofa-lookup form-text" id="lot-number" placeholder="Search by lot number" data-drupal-selector="edit-lot-number" type="text" name="lot_number" value="" size="60" maxlength="128" /> </div> <input data-drupal-selector="edit-brand" type="hidden" name="brand" value="novus" /> <input data-drupal-selector="edit-document-category" type="hidden" name="document_category" value="coa" /> </div> <div class="col-auto mb-3"> <input id="edit-submit" class="btn btn-primary cofa-btn button js-form-submit form-submit" data-drupal-selector="edit-submit" type="submit" name="op" value="Submit" /> </div> </div> </form> </div> <div id="pdf-result-novus" class="col-md-6 mb-4 pdf-result datasheet-loading"></div> </section> </form> </div> </div> </div> </div> <div id="specific-notices" class="py-5" data-section="section-base--specific-notices"> <h3>Product Specific Notices for CRISPR-Cas9 Antibody Pack </h3> <p></p><p>This product is for research use only and is not approved for use in humans or in clinical diagnosis. 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