

Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN and ChIP
{"AddThisServiceButtonMeta":"","add_this":[{"annotation":"","service":"facebook"},{"annotation":"","service":"email"},{"annotation":"","service":"print"},{"annotation":"","service":"twitter"},{"annotation":"","service":"linkedin"}],"add_to_wishlist_url":"/wishlist.php?action=add&product_id=760","availability":"","bulk_discount_rates":[],"can_purchase":true,"cart_url":"https://www.epicypher.com/cart.php","category":["Nucleosomes/SNAP-CUTANA™ Spike-in Controls","Antibodies","Antibodies/CUTANA™ CUT&RUN Antibodies","Antibodies/CUTANA™ CUT&RUN Antibodies/CUT&RUN Antibodies - Histone PTMs","Antibodies/SNAP-ChIP<sup>®</sup> Certified Antibodies","Epigenetics Kits and Reagents/CUTANA™ ChIC / CUT&RUN Assays","Epigenetics Kits and Reagents/CUTANA™ CUT&Tag Assays"],"custom_fields":[{"id":"684","name":"Pack Size","value":"100 µg"},{"id":"685","name":"Internal Comment","value":"extra box in top shelf of Venom"}],"customizations":[],"description":"<div class=\"product-general-info\">\n <ul class=\"product-general-info__list-left\">\n <li class=\"product-general-info__list-item\">\n <strong>Type: </strong>Mixed Monoclonal*\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Target Size: </strong>15 kDa\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Format: </strong>Affinity Purified IgG\n </li>\n </ul>\n <ul class=\"product-general-info__list-right\">\n <li class=\"product-general-info__list-item\">\n <strong>Host: </strong>Rabbit\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Reactivity: </strong>Human, Mouse, Drosophila, Yeast,\n Wide Range (Predicted)\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Applications: </strong>CUT&RUN, ChIP, ICC/IF, WB\n </li>\n </ul>\n</div>\n\n<div class=\"service_accordion product-droppdown\">\n <div class=\"container\">\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel current\">\n <h3 class=\"sub-title1\">Description</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description-specific\"\n >\n <p>\n This antibody meets EpiCypher's lot-specific\n SNAP-Certified™ criteria for specificity and efficient target\n enrichment in both CUT&RUN and ChIP applications. In CUT&RUN, this\n requires <20% cross reactivity to related histone PTMs\n determined using the SNAP-CUTANA™ K-MetStat Panel of spike-in\n controls (EpiCypher\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/snap-cutana-k-metstat-panel\"\n >19-1002</a\n >). High target efficiency is confirmed by consistent genomic\n enrichment at 500k and 50k starting cells. In ChIP, this requires\n <20% cross-reactivity to related histone PTMs and >5% of\n target input recovered as determined using the SNAP-ChIP<sup>®</sup>\n K-MetStat Panel of spike-in controls (EpiCypher\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/snap-chip-k-metstat-panel\"\n >19-1001</a\n >) [1].\n </p>\n\n <p>*Mixed Monoclonal: a pool of multiple monoclonal antibodies.</p>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel current\">\n <h3 class=\"sub-title1\">Validation Data</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description-specific\"\n >\n <section class=\"image-picker\">\n <div class=\"image-picker__left\">\n <div\n class=\"image-picker__main-content_active image-picker__main-content\"\n >\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-specificity-analysis.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"13-0041-specificity-analysis\"\n src=\"/content/images/products/antibodies/13-0041-specificity-analysis.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n ></a\n >\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\"\n ><strong>Figure 1: SNAP specificity analysis </strong><br />\n CUT&RUN was performed on 500k and 50k native K-562 cells\n with the SNAP-CUTANA™ K-MetStat Panel (EpiCypher\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/snap-cutana-k-metstat-panel\"\n >19-1002</a\n >) spiked-in prior to the addition of 0.5 µg H3K4me3\n antibody. CUT&RUN sequencing reads were aligned to the\n unique DNA barcodes corresponding to each nucleosome in the\n SNAP-CUTANA™ panel (x-axis). Data are expressed as a\n percent relative to on-target recovery (H3K4me3 set to\n 100%). Error bars represent mean ± SEM for the duplicate DNA\n barcodes in a single CUT&RUN experiment.\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-genome-wide-enrichment.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"13-0041-genome-wide-enrichment\"\n src=\"/content/images/products/antibodies/13-0041-genome-wide-enrichment.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n ></a\n >\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\"\n ><strong>Figure 2: CUT&RUN genome wide enrichment</strong\n ><br />\n CUT&RUN was performed as above (Figure 1). Sequence reads\n were aligned to 18,793 annotated transcription start sites\n (TSSs, ± 2 kbp). Signal enrichment was sorted from highest\n to lowest (top to bottom) relative to H3K4me3 - 50k sample\n (all gene rows aligned). High, medium, and low intensity are\n shown in red, yellow, and blue, respectively. H3K4me3\n antibody produced the expected TSS enrichment pattern that\n was consistent between 500k and 50k cells, while IgG\n negative control antibody (EpiCypher\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/snap-cutana-spike-in-controls/cutana-rabbit-igg-cut-run-negative-control-antibody\"\n >13-0042</a\n >) displayed minimal background.\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-representative-tracks.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"13-0041-representative-tracks\"\n src=\"/content/images/products/antibodies/13-0041-representative-tracks.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n >\n </a>\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\">\n <strong\n >Figure 3: H3K4me3 CUT&RUN representative tracks at 500k\n and 50k cells</strong\n ><br />\n Gene browser shots generated using the Integrative Genomics\n Viewer (IGV, Broad Institute) show representative loci for\n H3K4me3 antibody tested in CUT&RUN using 500k and 50k cells.\n Robust, specific enrichment is observed at gene\n transcription start sites for both cell inputs.\n <em\n ><u>Methods</u>: CUT&RUN was performed using the CUTANA™\n ChIC/CUT&RUN Kit v2.0 (EpiCypher\n <a\n href=\"/products/epigenetics-reagents-and-assays/cutana-chic-cut-and-run-kit\"\n >14-1048</a\n >) adapted to high throughput Tecan liquid handling.\n Library preparation was performed with 5 ng DNA using the\n NEBNext<sup>®</sup> Ultra™ II DNA Library Prep Kit for\n Illumina<sup>®</sup>. Libraries were run on an Illumina\n NextSeq2000 with paired-end sequencing (2x50 bp). Sample\n sequencing depth was 5.7 million reads (500k cell input)\n and 5.6 million reads (50k cell input).</em\n >\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-correlation-analysis.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"13-0041-correlation-analysis\"\n src=\"/content/images/products/antibodies/13-0041-correlation-analysis.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n >\n </a>\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\">\n <strong\n >Figure 4: CUT&RUN cell input correlation analysis </strong\n ><br />\n Genome-wide correlation analysis was performed to compare\n H3K4me3 antibody enrichment in CUT&RUN using 500k and 50k\n cell inputs. The log of the number of reads per 75 bp binned\n region across the genome is plotted for both samples.\n CUT&RUN data generated using this H3K4me3 antibody are\n highly correlated between the two cell inputs (Pearson\n correlation r = 0.828), indicating high consistency of\n H3K4me3 antibody target recovery.\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-multiplexed-specificity-profiling.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"13-0041-multiplexed-specificity-profiling\"\n src=\"/content/images/products/antibodies/13-0041-multiplexed-specificity-profiling.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n >\n </a>\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\">\n <strong>Figure 5: Multiplexed specificity profiling </strong\n ><br />\n Histone H3K4me3 antibody was assessed using a Luminex<sup\n >®</sup\n >\n based approach employing dCypher™ Nucleosome K-MetStat\n Panel (EpiCypher\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/dcypher-nucleosome-panels/dcypher-nucleosome-k-metstat-oncostat-panel\"\n >16-9002</a\n >). The panel comprises biotinylated designer nucleosomes\n (x-axis) individually coupled to color coded Luminex\n MagPlex<up>®</up> beads. Antibody binding to the panel of 16\n nucleosomes was tested in multiplex at a 1:250 dilution and\n detected with a second layer anti-IgG*PE. Data was generated\n using a Luminex FlexMAP 3D<sup>®</sup>. Data is normalized\n to relevant on-target (H3K4me3; set to 100).\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-immunofluorescence.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"13-0041-immunofluorescence\"\n src=\"/content/images/products/antibodies/13-0041-immunofluorescence.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n >\n </a>\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\">\n <strong>Figure 6: Immunofluorescence </strong><br />\n Representative images (60X magnification) of HeLa cells\n fixed, permeabilized, and immunostained to show endogenous\n localization of H3K4me3. <strong>Panel a:</strong> H3K4me3\n antibody (green, 1:100 dilution) detected with an Alexa\n Fluor<sup>®</sup> 488 conjugated anti-rabbit secondary.\n <strong>Panel b:</strong> DAPI stained nuclei (blue).\n <strong>Panel c:</strong> Rhodamine stained cytoskeletal\n F-actin (red). <strong>Panel d:</strong> A composite of\n panels a, b & c demonstrating the nuclear localization of\n H3K4me3. <strong>Panel e:</strong> Negative control lacking\n H3K4me3 primary antibody to assess background.\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-western-blot-data.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"13-0041-western-blot-data\"\n src=\"/content/images/products/antibodies/13-0041-western-blot-data.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n >\n </a>\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\">\n <strong>Figure 7: Western blot data </strong><br />\n Western analysis of H3K4me3 in whole cell extracts from\n HeLa, Hep G2, HCT 116, MCF7, U-2 OS, AF49, HEK-293, NIH/3T3,\n and PC-12 cells. 30 µg of lysate was resolved via SDS-PAGE\n and detected with H3K4me3 antibody at a 1:250 dilution.\n </span>\n </p>\n </div>\n <div class=\"image-picker__main-content\">\n <div class=\"image-picker__header-content\">\n <button class=\"image-picker__left-arrow\">\n <svg\n class=\"image-picker__svg-left\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M16.67 0l2.83 2.829-9.339 9.175 9.339 9.167-2.83 2.829-12.17-11.996z\"\n />\n </svg>\n </button>\n <a\n href=\"/content/images/products/antibodies/13-0041-specificity-enrichment-analysis.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"13-0041-specificity-enrichment-analysis\"\n src=\"/content/images/products/antibodies/13-0041-specificity-enrichment-analysis.jpeg\"\n class=\"image-picker__main-image\"\n />\n <span class=\"image-picker__main-image-caption\"\n >(Click to enlarge)</span\n >\n </a>\n <button class=\"image-picker__right-arrow\">\n <svg\n class=\"image-picker__svg-right\"\n width=\"24\"\n height=\"24\"\n viewBox=\"0 0 24 24\"\n >\n <path\n d=\"M7.33 24l-2.83-2.829 9.339-9.175-9.339-9.167 2.83-2.829 12.17 11.996z\"\n />\n </svg>\n </button>\n </div>\n <p>\n <span class=\"image-picker__span-content\">\n <strong\n >Figure 8: SNAP-ChIP-qPCR specificity and enrichment\n analysis </strong\n ><br />\n Histone H3K4me3 antibody (5 µg) was tested in a native ChIP\n experiment using chromatin from K-562 cells (5 µg) with the\n SNAP-ChIP<sup>®</sup> K-MetStat Panel (EpiCypher\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/snap-chip-k-metstat-panel\"\n >19-1001</a\n >) spiked-in prior to micrococcal nuclease digestion.\n Specificity (left y-axis) was determined by qPCR for the DNA\n barcodes corresponding to modified nucleosomes in the\n SNAP-ChIP<sup>®</sup>\n panel (x-axis). Black bar represents antibody efficiency\n (right y-axis; log scale) and indicates percentage of the\n target immunoprecipitated relative to input. Error bars\n represent mean ± SEM in replicate ChIP experiments.\n </span>\n </p>\n </div>\n </div>\n <aside class=\"image-picker__right\">\n <div class=\"image-picker__gallery\">\n <img\n alt=\"13-0041-specificity-analysis\"\n src=\"/content/images/products/antibodies/13-0041-specificity-analysis.jpeg\"\n width=\"200\"\n class=\"image-picker__side-image image-picker__side-image_active\"\n role=\"button\"\n />\n <img\n alt=\"13-0041-genome-wide-enrichment\"\n src=\"/content/images/products/antibodies/13-0041-genome-wide-enrichment.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"13-0041-representative-tracks\"\n src=\"/content/images/products/antibodies/13-0041-representative-tracks.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"13-0041-correlation-analysis\"\n src=\"/content/images/products/antibodies/13-0041-correlation-analysis.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"13-0041-multiplexed-specificity-profiling\"\n src=\"/content/images/products/antibodies/13-0041-multiplexed-specificity-profiling.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"13-0041-immunofluorescence\"\n src=\"/content/images/products/antibodies/13-0041-immunofluorescence.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"13-0041-western-blot-data\"\n src=\"/content/images/products/antibodies/13-0041-western-blot-data.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"13-0041-specificity-enrichment-analysis\"\n src=\"/content/images/products/antibodies/13-0041-specificity-enrichment-analysis.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n </div>\n </aside>\n </section>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Technical Information</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <div class=\"product-tech-info\">\n <div class=\"product-tech-info__line-item\">\n <div class=\"product-tech-info__line-item-left\">\n <b>Immunogen</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n A synthetic peptide corresponding to histone H3 trimethylated at\n lysine 4.\n </div>\n </div>\n <div class=\"product-tech-info__line-item\">\n <div class=\"product-tech-info__line-item-left\">\n <b>Formulation</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n Protein A affinity-purified antibody in PBS pH 7.4, with 0.09%\n sodium azide.\n </div>\n </div>\n <div class=\"product-tech-info__line-item\">\n <div class=\"product-tech-info__line-item-left\">\n <b>Storage and Stability</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n Stable for 1 year at -20°C from date of receipt.\n </div>\n </div>\n </div>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Recommended Dilution</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <p><strong>CUT&RUN:</strong> 0.5 µg per reaction</p>\n <p>\n <strong>Chromatin Immunoprecipitation (ChIP):</strong> 2 - 5 µg per\n 5 µg chromatin\n </p>\n <p>\n <strong>Immunocytochemistry/Immunofluorescence (ICC/IF):</strong>\n 1:100\n </p>\n <p><strong>Western Blot (WB):</strong> 1:250</p>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">References</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <strong>Background References:</strong>\n <br />\n [1] Shah et al. <em>Mol. Cell</em> (2018). PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/30244833/\"\n target=\"_blank\"\n title=\"Examining the Roles of H3K4 Methylation States with Systematically Characterized Antibodies\"\n >30244833</a\n ><br />\n <br />\n <strong>Product References:</strong>\n <iframe\n class=\"airtable-embed\"\n src=\"https://airtable.com/embed/shrk7LU4HS9DBrCv5?backgroundColor=purple&viewControls=on\"\n frameborder=\"0\"\n onmousewheel=\"\"\n width=\"100%\"\n height=\"300\"\n style=\"background: transparent; border: 1px solid #ccc\"\n ></iframe>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Documents & Resources</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <div class=\"product-documents\">\n <a\n href=\"/content/documents/tds/13-0041.pdf\"\n target=\"_blank\"\n class=\"product-documents__link\"\n >\n <svg\n version=\"1.1\"\n id=\"Layer_1\"\n xmlns=\"http://www.w3.org/2000/svg\"\n xmlns:xlink=\"http://www.w3.org/1999/xlink\"\n x=\"0px\"\n y=\"0px\"\n viewBox=\"0 0 228 240\"\n style=\"enable-background: new 0 0 228 240\"\n xml:space=\"preserve\"\n class=\"product-documents__icon\"\n alt=\"16-0030 Datasheet\"\n >\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M191.92,68.77l-47.69-47.69c-1.33-1.33-3.12-2.08-5.01-2.08H45.09C41.17,19,38,22.17,38,26.09v184.36\n c0,3.92,3.17,7.09,7.09,7.09h141.82c3.92,0,7.09-3.17,7.09-7.09V73.8C194,71.92,193.25,70.1,191.92,68.77z M177.65,77.06h-41.7\n v-41.7L177.65,77.06z M178.05,201.59H53.95V34.95h66.92v47.86c0,5.14,4.17,9.31,9.31,9.31h47.86V201.59z\"\n />\n </g>\n <rect\n x=\"20\"\n y=\"112\"\n class=\"product-documents__svg-background\"\n width=\"146\"\n height=\"76\"\n />\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M23.83,125.68h22.36c5.29,0,9.41,1.33,12.35,4c2.94,2.67,4.42,6.39,4.42,11.18c0,4.78-1.47,8.51-4.42,11.18\n c-2.94,2.67-7.06,4-12.35,4H34.59v18.29H23.83V125.68z M44.81,147.9c5.38,0,8.07-2.32,8.07-6.97c0-2.39-0.67-4.16-2-5.31\n c-1.33-1.15-3.36-1.73-6.07-1.73H34.59v14.01H44.81z\"\n />\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M69.92,125.68h18.91c5.29,0,9.84,0.97,13.66,2.9c3.82,1.93,6.74,4.72,8.76,8.35\n c2.02,3.63,3.04,7.98,3.04,13.04c0,5.06-1,9.42-3,13.08c-2,3.66-4.91,6.45-8.73,8.38c-3.82,1.93-8.4,2.9-13.73,2.9H69.92V125.68z\n M88.07,165.63c10.35,0,15.52-5.22,15.52-15.66c0-10.4-5.17-15.59-15.52-15.59h-7.38v31.26H88.07z\"\n />\n <path\n class=\"product-documents__svg-pdf\"\n d=\"M122.57,125.68h32.84v8.49h-22.22v11.18h20.84v8.49h-20.84v20.49h-10.63V125.68z\"\n />\n </g>\n </svg>\n <span class=\"product-documents__info\">Technical Datasheet</span>\n </a>\n </div>\n </div>\n </div>\n </div>\n </div>\n</div>\n","detail_messages":"","gift_wrapping_available":false,"gtin":null,"id":"760","images":[{"alt":"Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN and ChIP","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/760/743/snap-chip-ab__70507.1557259520.1280.1280__90586.1575483123.1280.1280__61997.1592491196.png?c=2"}],"main_image":{"alt":"Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN and ChIP","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/760/743/snap-chip-ab__70507.1557259520.1280.1280__90586.1575483123.1280.1280__61997.1592491196.png?c=2"},"max_purchase_quantity":0,"meta_description":"Rabbit mixed monoclonal histone H3K4me3 antibody rigorously tested for robust and reliable performance in CUT&RUN and ChIP assays. Also validated for ICC/IF & Western Blot.","meta_keywords":"H3K4me3 antibody, histone H3K4me3 antibody, histone H3 lysine 4 trimethylated antibody, ChIP antibody, CUT&RUN antibody","min_purchase_quantity":0,"mpn":null,"num_reviews":0,"options":[],"page_title":"H3K4me3 Antibody | SNAP-Certified for CUT&RUN and ChIP","price":{"tax_label":"Sales Tax","without_tax":{"currency":"USD","formatted":"$475.00","value":475}},"rating":0,"related_products":[{"add_to_cart_url":"https://www.epicypher.com/cart.php?action=add&product_id=872","add_to_wishlist_url":"/wishlist.php?action=add&product_id=872","availability":"","brand":{"name":null},"category":["Antibodies/SNAP-ChIP<sup>®</sup> Certified Antibodies"],"custom_fields":[{"id":893,"name":"Pack Size","value":"100 µL"}],"date_added":"19th Jul 2021","demo":false,"has_options":false,"id":872,"image":{"alt":"Histone H3K14ac Antibody, SNAP-ChIP® Certified","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/872/899/snap-chip-ab__70507.1557259520.1280.1280__90586.1575485337.190.250__92644.1626671512.png?c=2"},"images":[{"alt":"Histone H3K14ac Antibody, SNAP-ChIP® Certified","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/872/899/snap-chip-ab__70507.1557259520.1280.1280__90586.1575485337.190.250__92644.1626671512.png?c=2"}],"low_stock_level":null,"name":"Histone H3K14ac Antibody, SNAP-ChIP® Certified","num_reviews":null,"pre_order":false,"price":{"tax_label":"Sales Tax","without_tax":{"currency":"USD","formatted":"$465.00","value":465}},"qty_in_cart":0,"rating":null,"show_cart_action":true,"sku":"13-0049","stock_level":null,"summary":"\n \n \n Type: Recombinant Monoclonal\n \n \n Target Size: 15 kDa\n \n \n Format: Affinity Purified IgG\n \n \n \n \n ...","url":"https://www.epicypher.com/products/antibodies/snap-chip-sup-sup-certified-antibodies/histone-h3k14ac-antibody-snap-chip-certified","weight":{"formatted":"0.00 LBS","value":0}},{"add_to_cart_url":"https://www.epicypher.com/cart.php?action=add&product_id=892","add_to_wishlist_url":"/wishlist.php?action=add&product_id=892","availability":"","brand":{"name":null},"category":["Antibodies/CUTANA™ CUT&RUN Antibodies","Antibodies/CUTANA™ CUT&RUN Antibodies/CUT&RUN Antibodies - Histone PTMs"],"custom_fields":[{"id":988,"name":"Pack Size","value":"100 µg"}],"date_added":"17th Feb 2022","demo":false,"has_options":false,"id":892,"image":{"alt":"Histone H3K27me1 Antibody, SNAP-Certified for CUTandRUN","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/892/954/histone-h3k27me1-antibody-snap-certified-for-cutandrun__98289.1645734546.jpg?c=2"},"images":[{"alt":"Histone H3K27me1 Antibody, SNAP-Certified for CUTandRUN","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/892/954/histone-h3k27me1-antibody-snap-certified-for-cutandrun__98289.1645734546.jpg?c=2"}],"low_stock_level":null,"name":"Histone H3K27me1 Antibody, SNAP-Certified™ for CUT&RUN","num_reviews":null,"pre_order":false,"price":{"tax_label":"Sales Tax","without_tax":{"currency":"USD","formatted":"$475.00","value":475}},"qty_in_cart":0,"rating":null,"show_cart_action":true,"sku":"13-0052","stock_level":null,"summary":"\n \n \n Type: Mixed Monoclonal*\n \n \n Target Size: 15 kDa\n \n \n Format: Affinity Purified IgG\n \n \n \n \n ...","url":"https://www.epicypher.com/products/antibodies/cut-and-run-antibodies/cut-and-run-antibodies-histone-ptms/histone-h3k27me1-antibody-snap-certified-for-cut-and-run","weight":{"formatted":"0.01 LBS","value":0.01}},{"add_to_cart_url":"https://www.epicypher.com/cart.php?action=add&product_id=893","add_to_wishlist_url":"/wishlist.php?action=add&product_id=893","availability":"","brand":{"name":null},"category":["Antibodies/CUTANA™ CUT&RUN Antibodies","Antibodies/CUTANA™ CUT&RUN Antibodies/CUT&RUN Antibodies - Histone PTMs"],"custom_fields":[{"id":989,"name":"Pack Size","value":"100 µg"}],"date_added":"17th Feb 2022","demo":false,"has_options":false,"id":893,"image":{"alt":"Histone H4K20me3 Antibody, SNAP-Certified™ for CUT&RUN","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/893/923/1628650844.190.250__22739.1628695556__57006.1645069385.png?c=2"},"images":[{"alt":"Histone H4K20me3 Antibody, SNAP-Certified™ for CUT&RUN","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/893/923/1628650844.190.250__22739.1628695556__57006.1645069385.png?c=2"}],"low_stock_level":null,"name":"Histone H4K20me3 Antibody, SNAP-Certified™ for CUT&RUN","num_reviews":null,"pre_order":false,"price":{"tax_label":"Sales Tax","without_tax":{"currency":"USD","formatted":"$475.00","value":475}},"qty_in_cart":0,"rating":null,"show_cart_action":true,"sku":"13-0054","stock_level":null,"summary":"\n \n \n Type: Monoclonal\n \n \n Target Size: 15 kDa\n \n \n Format: Affinity Purified IgG\n \n \n \n \n Host: Mouse\n \n ...","url":"https://www.epicypher.com/products/antibodies/cut-and-run-antibodies/cut-and-run-antibodies-histone-ptms/histone-h4k20me3-antibody-snap-certified-for-cut-and-run","weight":{"formatted":"0.01 LBS","value":0.01}},{"add_to_wishlist_url":"/wishlist.php?action=add&product_id=626","availability":"","brand":{"name":null},"category":[],"custom_fields":[{"id":452,"name":"Pack Size","value":"100 μg"}],"date_added":"24th Sep 2018","demo":false,"has_options":false,"id":626,"image":{"alt":"Histone H3K4me3 Antibody, SNAP-ChIP Certified *DISCONTINUED*","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/626/578/Screen_Shot_2018-09-25_at_2.00.27_PM__52796.1538135192.png?c=2"},"images":[{"alt":"Histone H3K4me3 Antibody, SNAP-ChIP Certified *DISCONTINUED*","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/626/578/Screen_Shot_2018-09-25_at_2.00.27_PM__52796.1538135192.png?c=2"}],"low_stock_level":null,"name":"Histone H3K4me3 Antibody, SNAP-ChIP Certified *DISCONTINUED*","num_reviews":null,"pre_order":false,"price":{"tax_label":"Sales Tax","without_tax":{"currency":"USD","formatted":"$465.00","value":465}},"qty_in_cart":0,"rating":null,"show_cart_action":false,"sku":"13-0028","stock_level":null,"summary":"\n \n \n \n \n Product Discontinued - See 13-0041 as a recommended alternative\n \n \n \n \n \n \n \n \n ...","url":"https://www.epicypher.com/products/antibodies/snap-chip-certified-antibodies/histone-h3k4me3-antibody-snap-chip-certified","weight":{"formatted":"0.00 LBS","value":0}},{"add_to_cart_url":"https://www.epicypher.com/cart.php?action=add&product_id=627","add_to_wishlist_url":"/wishlist.php?action=add&product_id=627","availability":"","brand":{"name":null},"category":["Antibodies/CUTANA™ CUT&RUN Antibodies","Antibodies/CUTANA™ CUT&RUN Antibodies/CUT&RUN Antibodies - Histone PTMs","Antibodies/SNAP-ChIP<sup>®</sup> Certified Antibodies"],"custom_fields":[{"id":453,"name":"Pack Size","value":"100 μg"}],"date_added":"24th Sep 2018","demo":false,"has_options":false,"id":627,"image":{"alt":"Histone H3K9me1 Antibody, SNAP-Certified™ for CUT&RUN and ChIP","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/627/579/Screen_Shot_2018-09-25_at_2.00.27_PM__31915.1538135192.png?c=2"},"images":[{"alt":"Histone H3K9me1 Antibody, SNAP-Certified™ for CUT&RUN and ChIP","data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/627/579/Screen_Shot_2018-09-25_at_2.00.27_PM__31915.1538135192.png?c=2"}],"low_stock_level":null,"name":"Histone H3K9me1 Antibody, SNAP-Certified™ for CUT&RUN and ChIP","num_reviews":null,"pre_order":false,"price":{"tax_label":"Sales Tax","without_tax":{"currency":"USD","formatted":"$475.00","value":475}},"qty_in_cart":0,"rating":null,"show_cart_action":true,"sku":"13-0029","stock_level":null,"summary":"\n \n \n Type: Monoclonal\n \n \n Target Size: 15 kDa\n \n \n Format: Affinity Purified IgG\n \n \n \n \n Host: Rabbit\n \n ...","url":"https://www.epicypher.com/products/antibodies/cut-and-run-antibodies/cut-and-run-antibodies-histone-ptms/histone-h3k9me1-antibody-snap-certified-for-cut-run-and-chip","weight":{"formatted":"0.00 LBS","value":0}}],"shipping":{"calculated":true},"shipping_messages":[],"show_quantity_input":1,"sku":"13-0041","tags":[],"title":"Histone H3K4me3 Antibody, SNAP-Certified™ for CUT&RUN and ChIP","upc":null,"url":"https://www.epicypher.com/products/antibodies/snap-chip-certified-antibodies/histone-h3k4me3-antibody-snap-chip-certified-cutana-cut-run-compatible","warranty":"","weight":"0.01 LBS"} Pack Size: 100 µg
- Type: Mixed Monoclonal*
- Target Size: 15 kDa
- Format: Affinity Purified IgG
- Host: Rabbit
- Reactivity: Human, Mouse, Drosophila, Yeast, Wide Range (Predicted)
- Applications: CUT&RUN, ChIP, ICC/IF, WB
Description
This antibody meets EpiCypher's lot-specific SNAP-Certified™ criteria for specificity and efficient target enrichment in both CUT&RUN and ChIP applications. In CUT&RUN, this requires <20% cross reactivity to related histone PTMs determined using the SNAP-CUTANA™ K-MetStat Panel of spike-in controls (EpiCypher 19-1002). High target efficiency is confirmed by consistent genomic enrichment at 500k and 50k starting cells. In ChIP, this requires <20% cross-reactivity to related histone PTMs and >5% of target input recovered as determined using the SNAP-ChIP® K-MetStat Panel of spike-in controls (EpiCypher 19-1001) [1].
*Mixed Monoclonal: a pool of multiple monoclonal antibodies.
Validation Data
Figure 1: SNAP specificity analysis
CUT&RUN was performed on 500k and 50k native K-562 cells
with the SNAP-CUTANA™ K-MetStat Panel (EpiCypher
19-1002) spiked-in prior to the addition of 0.5 µg H3K4me3
antibody. CUT&RUN sequencing reads were aligned to the
unique DNA barcodes corresponding to each nucleosome in the
SNAP-CUTANA™ panel (x-axis). Data are expressed as a
percent relative to on-target recovery (H3K4me3 set to
100%). Error bars represent mean ± SEM for the duplicate DNA
barcodes in a single CUT&RUN experiment.
Figure 2: CUT&RUN genome wide enrichment
CUT&RUN was performed as above (Figure 1). Sequence reads
were aligned to 18,793 annotated transcription start sites
(TSSs, ± 2 kbp). Signal enrichment was sorted from highest
to lowest (top to bottom) relative to H3K4me3 - 50k sample
(all gene rows aligned). High, medium, and low intensity are
shown in red, yellow, and blue, respectively. H3K4me3
antibody produced the expected TSS enrichment pattern that
was consistent between 500k and 50k cells, while IgG
negative control antibody (EpiCypher
13-0042) displayed minimal background.
Figure 3: H3K4me3 CUT&RUN representative tracks at 500k
and 50k cells
Gene browser shots generated using the Integrative Genomics
Viewer (IGV, Broad Institute) show representative loci for
H3K4me3 antibody tested in CUT&RUN using 500k and 50k cells.
Robust, specific enrichment is observed at gene
transcription start sites for both cell inputs.
Methods: CUT&RUN was performed using the CUTANA™
ChIC/CUT&RUN Kit v2.0 (EpiCypher
14-1048) adapted to high throughput Tecan liquid handling.
Library preparation was performed with 5 ng DNA using the
NEBNext® Ultra™ II DNA Library Prep Kit for
Illumina®. Libraries were run on an Illumina
NextSeq2000 with paired-end sequencing (2x50 bp). Sample
sequencing depth was 5.7 million reads (500k cell input)
and 5.6 million reads (50k cell input).
Figure 4: CUT&RUN cell input correlation analysis
Genome-wide correlation analysis was performed to compare
H3K4me3 antibody enrichment in CUT&RUN using 500k and 50k
cell inputs. The log of the number of reads per 75 bp binned
region across the genome is plotted for both samples.
CUT&RUN data generated using this H3K4me3 antibody are
highly correlated between the two cell inputs (Pearson
correlation r = 0.828), indicating high consistency of
H3K4me3 antibody target recovery.
Figure 5: Multiplexed specificity profiling
Histone H3K4me3 antibody was assessed using a Luminex®
based approach employing dCypher™ Nucleosome K-MetStat
Panel (EpiCypher
16-9002). The panel comprises biotinylated designer nucleosomes
(x-axis) individually coupled to color coded Luminex
MagPlex
Figure 6: Immunofluorescence
Representative images (60X magnification) of HeLa cells
fixed, permeabilized, and immunostained to show endogenous
localization of H3K4me3. Panel a: H3K4me3
antibody (green, 1:100 dilution) detected with an Alexa
Fluor® 488 conjugated anti-rabbit secondary.
Panel b: DAPI stained nuclei (blue).
Panel c: Rhodamine stained cytoskeletal
F-actin (red). Panel d: A composite of
panels a, b & c demonstrating the nuclear localization of
H3K4me3. Panel e: Negative control lacking
H3K4me3 primary antibody to assess background.
Figure 7: Western blot data
Western analysis of H3K4me3 in whole cell extracts from
HeLa, Hep G2, HCT 116, MCF7, U-2 OS, AF49, HEK-293, NIH/3T3,
and PC-12 cells. 30 µg of lysate was resolved via SDS-PAGE
and detected with H3K4me3 antibody at a 1:250 dilution.
Figure 8: SNAP-ChIP-qPCR specificity and enrichment
analysis
Histone H3K4me3 antibody (5 µg) was tested in a native ChIP
experiment using chromatin from K-562 cells (5 µg) with the
SNAP-ChIP® K-MetStat Panel (EpiCypher
19-1001) spiked-in prior to micrococcal nuclease digestion.
Specificity (left y-axis) was determined by qPCR for the DNA
barcodes corresponding to modified nucleosomes in the
SNAP-ChIP®
panel (x-axis). Black bar represents antibody efficiency
(right y-axis; log scale) and indicates percentage of the
target immunoprecipitated relative to input. Error bars
represent mean ± SEM in replicate ChIP experiments.
Technical Information
Recommended Dilution
CUT&RUN: 0.5 µg per reaction
Chromatin Immunoprecipitation (ChIP): 2 - 5 µg per 5 µg chromatin
Immunocytochemistry/Immunofluorescence (ICC/IF): 1:100
Western Blot (WB): 1:250
References
Documents & Resources
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