Nucleosome, Recombinant Human, Acidic Patch Mutant H2BE105A,E113A
{"url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2be105a-e113a","add_this":[{"service":"facebook","annotation":""},{"service":"email","annotation":""},{"service":"print","annotation":""},{"service":"twitter","annotation":""},{"service":"linkedin","annotation":""}],"gtin":null,"id":866,"bulk_discount_rates":[],"can_purchase":true,"meta_description":"Acidic patch mutant recombinant mononucleosomes (H2BE105A,E113A), 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/866/956/nucleosome-recombinant-human-acidic-patch-mutant-h2be105ae113a__31522.1645734548.jpg?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2BE105A,E113A"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=866","shipping":{"calculated":true},"num_reviews":0,"weight":"0.00 LBS","custom_fields":[{"id":"801","name":"Pack Size","value":"50 µg"}],"sku":"16-1031","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 H2B contains a glutamate-to-alanine (E-to-A) substitution\n at positions 105 and 113 (H2BE105A,E113A). H2BE105 and H2BE113 are\n among key residues forming a negatively charged region on the\n nucleosome surface named the “acidic patch”. The acidic patch is a\n conserved interaction hub for neighboring nucleosomes and nucleosome\n binding proteins, often via salt bridges with arginine anchors, and\n is functionally critical in chromatin condensation and chromatin\n remodeling [2-4]. H2B E105 and E113 both reside in the H2B alphaC\n extension and are associated with nucleosome binding factors such as\n histone H4 N-terminal tail, LANA, RCC1, HMGN2 and SMARCB1 [2,5].\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/16-1031-protein-gel-data.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"16-1031-protein-gel-data\"\n src=\"/content/images/products/nucleosomes/16-1031-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 H2BE105A,E113A\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 (H2A,\n H2BE105A,E113A, 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/16-1031-dna-gel-data.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"16-1031-dna-gel-data\"\n src=\"/content/images/products/nucleosomes/16-1031-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 H2BE105A,E113A mononucleosomes resolved by native PAGE and\n stained 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/16-1031-functional-binding-assay.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"16-1031-functional-binding-assay\"\n src=\"/content/images/products/nucleosomes/16-1031-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 acidic patch mutations disrupts LANA peptide\n 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 >; H2AE92K, EpiCypher\n <a\n href=\"/products/nucleosomes/acidic-patch-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2ae92k\"\n >16-1030</a\n >; H2BE105A,E113A, EpiCypher 16-1031). The binding of\n biotinylated LANA peptide to recombinant nucleosomes was\n assessed by AlphaLISA assay (Perkin Elmer) using\n Streptavidin Donor Beads, anti-Histone H3.1/3.2 antibody,\n and Protein A 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=\"16-1031-protein-gel-data\"\n src=\"/content/images/products/nucleosomes/16-1031-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=\"16-1031_dna_gel-data\"\n src=\"/content/images/products/nucleosomes/16-1031-dna-gel-data.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"16-1031-functional-binding-assay\"\n src=\"/content/images/products/nucleosomes/16-1031-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 H2BE105A,E113A 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 = 199,028.7 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 H2BE105A,E113A mononucleosomes are highly purified and suitable for\n a variety of applications to test the effect of acidic patch\n mutations on enzymatic activity or chromatin binding. See EpiCypher\n <a\n href=\"/products/nucleosomes/acidic-patch-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2be105a-e113a-biotinylated\"\n >16-0031</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 (5) Valencia <em>et al.</em> <em>Cell</em> (2019). PMID:\n <a\n href=\"https://pubmed.ncbi.nlm.nih.gov/31759698/\"\n target=\"_blank\"\n title=\"Recurrent SMARCB1 Mutations Reveal a Nucleosome Acidic Patch Interaction Site That Potentiates mSWI/SNF Complex Chromatin Remodeling\"\n >31759698</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/16-1031.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=\"16-1031 Datasheet\"\n >\n <g>\n <path\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 | H2BE105A,E113A","cart_url":"https://www.epicypher.com/cart.php","max_purchase_quantity":0,"mpn":null,"upc":null,"options":[],"related_products":[{"id":861,"sku":"16-0031","name":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2BE105A,E113A Biotinylated","url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2be105a-e113a-biotinylated","availability":"","rating":null,"brand":{"name":null},"category":["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/861/927/nucleosome-recombinant-human-acidic-patch-mutant-h2be105ae113a-biotinylated__42717.1645734518.jpg?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2BE105A,E113A Biotinylated"},"images":[{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/861/927/nucleosome-recombinant-human-acidic-patch-mutant-h2be105ae113a-biotinylated__42717.1645734518.jpg?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2BE105A,E113A Biotinylated"},{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/861/885/2021.05.05_acidic_patch_nuc_RGB_LS__97220.1621279281.png?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2BE105A,E113A Biotinylated"}],"date_added":"17th May 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":806,"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=861","price":{"without_tax":{"currency":"USD","formatted":"$475.00","value":475},"tax_label":"Sales Tax"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=861"},{"id":1147,"sku":"16-2031","name":"Acidic Patch Mutant H2BE105A,E113A Recombinant Nucleosome with Linker DNA, Biotinylated","url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/acidic-patch-mutant-h2be105a-e113a-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/1147/1219/2021.05.05_acidic_patch_nuc_RGB_white_border_LS_icon_thumbnail_board_2__84943.1716971267.png?c=2","alt":"Acidic Patch Mutant H2BE105A,E113A Recombinant Nucleosome with Linker DNA, Biotinylated"},"images":[{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/1147/1219/2021.05.05_acidic_patch_nuc_RGB_white_border_LS_icon_thumbnail_board_2__84943.1716971267.png?c=2","alt":"Acidic Patch Mutant H2BE105A,E113A Recombinant Nucleosome with Linker DNA, Biotinylated"}],"date_added":"29th 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":1314,"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=1147","price":{"without_tax":{"currency":"USD","formatted":"$475.00","value":475},"tax_label":"Sales Tax"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=1147"},{"id":865,"sku":"16-1030","name":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE92K","url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2ae92k","availability":"","rating":null,"brand":{"name":null},"category":["Nucleosomes/Mutant Nucleosomes","Nucleosomes/Mutant Nucleosomes/Acidic Patch Nucleosomes"],"summary":"\n \n \n \n Description\n \n \n Mono","image":{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/865/890/2021.05.05_acidic_patch_nuc_RGB_white_border_LS_icon_thumbnail_board_2__87356.1626114896.png?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE92K"},"images":[{"data":"https://cdn11.bigcommerce.com/s-y9o92/images/stencil/{:size}/products/865/890/2021.05.05_acidic_patch_nuc_RGB_white_border_LS_icon_thumbnail_board_2__87356.1626114896.png?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE92K"}],"date_added":"23rd May 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":802,"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=865","price":{"without_tax":{"currency":"USD","formatted":"$475.00","value":475},"tax_label":"Sales Tax"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=865"},{"id":864,"sku":"16-1029","name":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE61A","url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2ae61a","availability":"","rating":null,"brand":{"name":null},"category":["Nucleosomes/Mutant Nucleosomes","Nucleosomes/Mutant Nucleosomes/Acidic Patch Nucleosomes"],"summary":"\n \n \n \n Description\n \n \n Mono","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"},"images":[{"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 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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 H2B contains a glutamate-to-alanine (E-to-A) substitution at positions 105 and 113 (H2BE105A,E113A). H2BE105 and H2BE113 are 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]. H2B E105 and E113 both reside in the H2B alphaC extension and are associated with nucleosome binding factors such as histone H4 N-terminal tail, LANA, RCC1, HMGN2 and SMARCB1 [2,5].
Validation Data
Figure 1: Protein Gel Data
Coomassie stained PAGE gel of proteins in H2BE105A,E113A
mononucleosomes (1 µg) demonstrates the purity of the
histones in the preparation. Sizes of molecular weight
markers and positions of the core histones (H2A,
H2BE105A,E113A, H3.1 and H4) are indicated.
Figure 2: DNA Gel Data
H2BE105A,E113A 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 acidic patch mutations disrupts LANA peptide
binding to recombinant nucleosomes (WT rNuc control,
EpiCypher
16-0009; H2AE92K, EpiCypher
16-1030; H2BE105A,E113A, EpiCypher 16-1031). 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
H2BE105A,E113A mononucleosomes are highly purified and suitable for a variety of applications to test the effect of acidic patch mutations on enzymatic activity or chromatin binding. See EpiCypher 16-0031 for a biotinylated version of this mutant.