Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE61A
{"url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2ae61a","add_this":[{"service":"facebook","annotation":""},{"service":"email","annotation":""},{"service":"print","annotation":""},{"service":"twitter","annotation":""},{"service":"linkedin","annotation":""}],"gtin":null,"id":864,"bulk_discount_rates":[],"can_purchase":true,"meta_description":"Acidic patch mutant recombinant mononucleosomes (H2AE61A), functionally validated with exceptional quality controls. Useful for chromatin binding, enzymatic, and structural studies.","category":["Nucleosomes/Mutant Nucleosomes","Nucleosomes/Mutant Nucleosomes/Acidic Patch Nucleosomes"],"AddThisServiceButtonMeta":"","main_image":{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/864/961/nucleosome-recombinant-human-acidic-patch-mutant-h2ae61a__21865.1645734553.jpg?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE61A"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=864","shipping":{"calculated":true},"num_reviews":0,"weight":"0.00 LBS","custom_fields":[{"id":"803","name":"Pack Size","value":"50 µg"}],"sku":"16-1029","description":"<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=\"\n ProductDescriptionContainer\n product-droppdown__section-description-specific\n \"\n >\n <p>\n Mononucleosomes assembled from recombinant human histones expressed\n in <em>E. coli</em> (two each of histones H2A*, H2B, H3 and H4;\n accession numbers: H2A-P04908*; H2B-O60814; H3.1-P68431; H4-P62805)\n wrapped by 147 base pairs of 601 positioning sequence DNA [1].\n </p>\n <p>\n *Histone H2A contains a glutamate-to-alanine (E-to-A) substitution\n at position 61 (H2AE61A). H2AE61A is among key residues forming a\n negatively charged region on the nucleosome surface named the\n “acidic patch”. The acidic patch is a conserved interaction hub for\n neighboring nucleosomes and nucleosome binding proteins, often via\n salt bridges with arginine anchors, and is functionally critical in\n chromatin condensation and chromatin remodeling [2-4]. H2AE61\n resides in the alpha2 helix and forms a key salt bridge with H4K16.\n H2AE61 mediates chromatin binding with factors such as LANA, RCC1,\n IL-33, SIR3 and HMGN2 [2]. H2AE61A disrupts chromatin remodeling by\n the ISWI remodeler SNF2h [4].\n </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=\"\n ProductDescriptionContainer\n product-droppdown__section-description-specific\n \"\n >\n <section class=\"image-picker\">\n <div class=\"image-picker__left\">\n <div\n class=\"\n image-picker__main-content_active image-picker__main-content\n \"\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/nucleosomes/16-1029-protein-gel-data.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"16-1029-protein-gel-data\"\n src=\"/content/images/products/nucleosomes/16-1029-protein-gel-data.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: Protein Gel Data </strong><br />\n Coomassie stained PAGE gel of proteins in H2AE61A\n mononucleosomes (1 µg) demonstrates the purity of the\n histones in the preparation. Sizes of molecular weight\n markers and positions of the core histones (H2AE61A, H2B,\n H3.1 and H4) are indicated.\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/nucleosomes/16-1029-dna-gel-data.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"16-1029-dna-gel-data\"\n src=\"/content/images/products/nucleosomes/16-1029-dna-gel-data.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: DNA Gel Data </strong><br />\n H2AE61A mononucleosomes resolved by native PAGE and stained\n with ethidium bromide to visualize DNA.\n <strong>Lane 1:</strong> Free DNA (100 ng).\n <strong>Lane 2:</strong> Intact nucleosomes (400 ng).\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/nucleosomes/16-1029-functional-binding-assay.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"16-1029-functional-binding-assay\"\n src=\"/content/images/products/nucleosomes/16-1029-functional-binding-assay.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 3: Functional Binding Assay </strong><br />\n The presence of the acidic patch mutation disrupts LANA\n peptide binding to recombinant nucleosomes (WT rNuc control,\n EpiCypher\n <a\n href=\"https://www.epicypher.com/products/nucleosomes/mononucleosomes-recombinant-human\"\n >16-0009</a\n >; H2AE61A, EpiCypher 16-1029). The binding of biotinylated\n LANA peptide to recombinant nucleosomes was assessed by\n AlphaLISA assay (Perkin Elmer) using Streptavidin Donor\n Beads, anti-Histone H3.1/3.2 antibody, and Protein A\n Acceptor Beads.\n </span>\n </p>\n </div>\n </div>\n <aside class=\"image-picker__right\">\n <div class=\"image-picker__gallery\">\n <img\n alt=\"16-1029-protein-gel-data\"\n src=\"/content/images/products/nucleosomes/16-1029-protein-gel-data.jpeg\"\n width=\"200\"\n class=\"\n image-picker__side-image image-picker__side-image_active\n \"\n role=\"button\"\n />\n <img\n alt=\"16-1029_dna_gel-data\"\n src=\"/content/images/products/nucleosomes/16-1029-dna-gel-data.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"16-1029-functional-binding-assay\"\n src=\"/content/images/products/nucleosomes/16-1029-functional-binding-assay.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=\"\n ProductDescriptionContainer\n product-droppdown__section-description\n \"\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>Formulation</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n H2AE61A mononucleosomes (27.3 μg protein weight, 50 μg\n DNA+protein) in 10 mM Tris HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2\n mM DTT, 20% glycerol. MW = 199,144.9 Da.\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 six (6) months at -80°C from date of receipt. For\n best results, aliquot and avoid multiple freeze/thaws.\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\">Application Notes</h3>\n <div\n class=\"\n ProductDescriptionContainer\n product-droppdown__section-description\n \"\n >\n <p>\n H2AE61A mononucleosomes are highly purified and suitable for a\n variety of applications to test the effect of acidic patch mutation\n on enzymatic activity or chromatin binding. See EpiCypher\n <a\n href=\"/products/nucleosomes/acidic-patch-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2ae61a-biotinylated\"\n >16-0029</a\n >\n for a biotinylated version of this mutant.\n </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=\"\n ProductDescriptionContainer\n product-droppdown__section-description\n \"\n >\n <strong>Background References:</strong>\n <br />\n (1) Lowary & Widom <em>J. Mol. Biol.</em> (1998). PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/9514715/\"\n target=\"_blank\"\n title=\"New DNA sequence rules for high affinity binding to histone octamer and sequence-directed nucleosome positioning\"\n >9514715</a\n ><br />\n (2) Kalashnikova <em>et al.</em> <em>J. R. Soc. Interface</em> (2013).\n PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/23446052/\"\n target=\"_blank\"\n title=\"The role of the nucleosome acidic patch in modulating higher order chromatin structure\"\n >23446052</a\n ><br />\n (3) Levendosky & Bowman <em>eLife</em> (2019). PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/31094676/\"\n target=\"_blank\"\n title=\"Asymmetry between the two acidic patches dictates the direction of nucleosome sliding by the ISWI chromatin remodeler\"\n >31094676</a\n ><br />\n (4) Gamarra <em>et al.</em> <em>eLife</em> (2018). PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/29664398/\"\n target=\"_blank\"\n title=\"The nucleosomal acidic patch relieves auto-inhibition by the ISWI remodeler SNF2h\"\n >29664398</a\n ><br />\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=\"\n ProductDescriptionContainer\n product-droppdown__section-description\n \"\n >\n <div class=\"product-documents\">\n <a\n href=\"/content/documents/tds/16-1029.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=\"16-1029 Datasheet\"\n >\n <g>\n <path\n class=\"product-documents__svg-pdf\"\n 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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","tags":[],"warranty":"","price":{"without_tax":{"formatted":"$475.00","value":475,"currency":"USD"},"tax_label":"Sales Tax"},"detail_messages":"","availability":"","page_title":"Acidic Patch Mutant Recombinant Nucleosome | H2AE61A","cart_url":"https://www.epicypher.com/cart.php","max_purchase_quantity":0,"mpn":null,"upc":null,"options":[],"related_products":[{"id":863,"sku":"16-0029","name":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE61A 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2021","pre_order":false,"show_cart_action":true,"has_options":false,"stock_level":null,"low_stock_level":null,"qty_in_cart":0,"custom_fields":[{"id":804,"name":"Pack Size","value":"50 µg"}],"num_reviews":null,"weight":{"formatted":"0.00 LBS","value":0},"demo":false,"add_to_cart_url":"https://www.epicypher.com/cart.php?action=add&product_id=863","price":{"without_tax":{"currency":"USD","formatted":"$475.00","value":475},"tax_label":"Sales Tax"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=863"},{"id":1148,"sku":"16-2029","name":"Acidic Patch Mutant H2AE61A Recombinant Nucleosome with Linker DNA, Biotinylated","url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/acidic-patch-mutant-h2ae61a-recombinant-nucleosome-with-linker-dna-biotinylated","availability":"","rating":null,"brand":{"name":null},"category":["Nucleosomes","Nucleosomes/Mutant Nucleosomes","Nucleosomes/Mutant Nucleosomes/Acidic Patch Nucleosomes"],"summary":"\n \n \n Species: Human\n \n \n Source: E. coli & synthetic DNA\n \n \n \n ","image":{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/1148/1221/2021.05.05_acidic_patch_nuc_RGB_white_border_LS_icon_thumbnail_board_2__41959.1717092961.png?c=2","alt":"Acidic Patch Mutant H2AE61A Recombinant Nucleosome with Linker DNA, Biotinylated"},"images":[{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/1148/1221/2021.05.05_acidic_patch_nuc_RGB_white_border_LS_icon_thumbnail_board_2__41959.1717092961.png?c=2","alt":"Acidic Patch Mutant H2AE61A Recombinant Nucleosome with Linker DNA, Biotinylated"}],"date_added":"30th May 2024","pre_order":false,"show_cart_action":true,"has_options":false,"stock_level":null,"low_stock_level":null,"qty_in_cart":0,"custom_fields":[{"id":1315,"name":"Pack Size","value":"50 µg"}],"num_reviews":null,"weight":{"formatted":"0.00 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Description
Mononucleosomes assembled from recombinant human histones expressed in E. coli (two each of histones H2A*, H2B, H3 and H4; accession numbers: H2A-P04908*; H2B-O60814; H3.1-P68431; H4-P62805) wrapped by 147 base pairs of 601 positioning sequence DNA [1].
*Histone H2A contains a glutamate-to-alanine (E-to-A) substitution at position 61 (H2AE61A). H2AE61A is among key residues forming a negatively charged region on the nucleosome surface named the “acidic patch”. The acidic patch is a conserved interaction hub for neighboring nucleosomes and nucleosome binding proteins, often via salt bridges with arginine anchors, and is functionally critical in chromatin condensation and chromatin remodeling [2-4]. H2AE61 resides in the alpha2 helix and forms a key salt bridge with H4K16. H2AE61 mediates chromatin binding with factors such as LANA, RCC1, IL-33, SIR3 and HMGN2 [2]. H2AE61A disrupts chromatin remodeling by the ISWI remodeler SNF2h [4].
Validation Data
Figure 1: Protein Gel Data
Coomassie stained PAGE gel of proteins in H2AE61A
mononucleosomes (1 µg) demonstrates the purity of the
histones in the preparation. Sizes of molecular weight
markers and positions of the core histones (H2AE61A, H2B,
H3.1 and H4) are indicated.
Figure 2: DNA Gel Data
H2AE61A mononucleosomes resolved by native PAGE and stained
with ethidium bromide to visualize DNA.
Lane 1: Free DNA (100 ng).
Lane 2: Intact nucleosomes (400 ng).
Figure 3: Functional Binding Assay
The presence of the acidic patch mutation disrupts LANA
peptide binding to recombinant nucleosomes (WT rNuc control,
EpiCypher
16-0009; H2AE61A, EpiCypher 16-1029). The binding of biotinylated
LANA peptide to recombinant nucleosomes was assessed by
AlphaLISA assay (Perkin Elmer) using Streptavidin Donor
Beads, anti-Histone H3.1/3.2 antibody, and Protein A
Acceptor Beads.
Technical Information
Application Notes
H2AE61A mononucleosomes are highly purified and suitable for a variety of applications to test the effect of acidic patch mutation on enzymatic activity or chromatin binding. See EpiCypher 16-0029 for a biotinylated version of this mutant.