Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE61A Biotinylated
{"url":"https://www.epicypher.com/products/nucleosomes/mutant-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2ae61a-biotinylated","add_this":[{"service":"facebook","annotation":""},{"service":"email","annotation":""},{"service":"print","annotation":""},{"service":"twitter","annotation":""},{"service":"linkedin","annotation":""}],"gtin":null,"id":863,"bulk_discount_rates":[],"can_purchase":true,"meta_description":"Acidic patch mutant recombinant mononucleosomes (biotinylated 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/863/888/2021.05.05_acidic_patch_nuc_RGB_white_border_LS_icon_thumbnail_board_2__67572.1621809858.png?c=2","alt":"Nucleosome, Recombinant Human, Acidic Patch Mutant H2AE61A Biotinylated"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=863","shipping":{"calculated":true},"num_reviews":0,"weight":"0.00 LBS","custom_fields":[{"id":"804","name":"Pack Size","value":"50 µg"}],"sku":"16-0029","description":"<div class=\"product-general-info\">\n <ul class=\"product-general-info__list-left\">\n <li class=\"product-general-info__list-item\">\n <strong>Species: </strong>Human\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Source: </strong><em>E. coli</em> & synthetic DNA\n </li>\n </ul>\n <ul class=\"product-general-info__list-right\">\n <li class=\"product-general-info__list-item\">\n <strong>Tag: </strong>Biotinylated\n </li>\n <li class=\"product-general-info__list-item\">\n <strong>Molecular Weight: </strong>199,626 Da\n </li>\n </ul>\n</div>\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 Recombinant mononucleosomes consist of 147 base pairs of DNA wrapped\n around an octamer core of histone proteins (two each of H2A, H2B,\n H3.1, and H4) to form a nucleosome, the basic repeating unit of\n chromatin. The 147 bp 601 sequence, which contains a 5’ biotin-TEG\n group, has high affinity for histone octamers and is useful for\n nucleosome assembly [1]. Histone H2A contains a glutamate-to-alanine\n (E-to-A) substitution at position 61 (H2AE61A). H2AE61 is among key\n residues forming a negatively charged region on the nucleosome\n surface named the “acidic patch”. The acidic patch is a conserved\n interaction hub for neighboring nucleosomes and nucleosome binding\n proteins, often via salt bridges with arginine anchors, and is\n functionally critical in chromatin condensation and chromatin\n remodeling [2-4]. H2AE61 resides in the alpha2 helix and forms a key\n salt bridge with H4K16. H2AE61 mediates chromatin binding with\n factors such as LANA, RCC1, IL-33, SIR3, and HMGN2 [2]. H2AE61A\n disrupts chromatin remodeling by the ISWI remodeler SMARCA5/SNF2h\n [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=\"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/nucleosomes/16-0029-test.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"16-0029-protein-gel-data-test\"\n src=\"/content/images/products/nucleosomes/16-0029-protein-gel-data-test.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 gata </strong><br />\n Coomassie stained PAGE gel of proteins in H2AE61A\n mononucleosome (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, H3\n 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/16-0029-dna-gel-data-test.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n ><img\n alt=\"16-0029-dna-gel-data-test\"\n src=\"/content/images/products/nucleosomes/16-0029-dna-gel-data-test.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 mononucleosome resolved by native PAGE and stained\n with ethidium bromide to visualize DNA.\n <strong>Lane 1:</strong> Free DNA (EpiCypher\n <a\n href=\"/products/nucleosomes/nucleosome-assembly-601-sequence-dna-biotinylated\"\n target=\"_blank\"\n >18-0005</a\n >; 100 ng). <strong>Lane 2:</strong> Intact H2AE61A\n mononucleosomes (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/16-0029-functional-binding-assay-test.jpeg\"\n target=\"_blank\"\n class=\"image-picker__main-image-link\"\n >\n <img\n alt=\"16-0029-functional-binding-assay-test\"\n src=\"/content/images/products/nucleosomes/16-0029-functional-binding-assay-test.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 control, EpiCypher\n <a\n href=\"/products/nucleosomes/mononucleosomes-recombinant-human-biotinylated\"\n >16-0006</a\n >; H2AE61A, EpiCypher 16-0029; H2AE92K, EpiCypher\n <a\n href=\"/products/nucleosomes/acidic-patch-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2ae92k-biotinylated\"\n >16-0030</a\n >; H2BE105A,E113A, EpiCypher\n <a\n href=\"/products/nucleosomes/acidic-patch-nucleosomes/nucleosome-recombinant-human-acidic-patch-mutant-h2be105a-e113a-biotinylated\"\n >16-0031</a\n >). The binding of GST-tagged LANA peptide to biotinylated\n recombinant nucleosomes was assessed by AlphaLISA assay\n using Streptavidin Donor Beads and Glutathione Acceptor\n Beads (PerkinElmer). The presence of H2A acidic patch\n mutations completely blocks LANA binding, while H2B\n mutations cause a decrease in LANA binding affinity.\n </span>\n </p>\n </div>\n </div>\n <aside class=\"image-picker__right\">\n <div class=\"image-picker__gallery\">\n <img\n alt=\"16-0029-protein-gel-data-test\"\n src=\"/content/images/products/nucleosomes/16-0029-protein-gel-data-test.jpeg\"\n width=\"200\"\n class=\"image-picker__side-image image-picker__side-image_active\"\n role=\"button\"\n />\n <img\n alt=\"16-0029-dna-gel-data-test\"\n src=\"/content/images/products/nucleosomes/16-0029-dna-gel-data-test.jpeg\"\n class=\"image-picker__side-image\"\n role=\"button\"\n />\n <img\n alt=\"16-0029-functional-binding-assay-test\"\n src=\"/content/images/products/nucleosomes/16-0029-functional-binding-assay-test.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>Storage</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n Stable for six months at -80°C from date of receipt. For best\n results, aliquot and avoid freeze/thaws.\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 10 mM Tris-HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20%\n glycerol. (27.3 μg protein, 50 μg DNA + protein)\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=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <p>\n H2AE61A mononucleosome is highly purified and suitable for a variety\n of applications to test the effect of acidic patch mutation on\n enzymatic activity or chromatin binding. The biotinylated DNA\n enables affinity binding applications.\n </p>\n </div>\n </div>\n </div>\n <div id=\"prodAccordion\">\n <div id=\"ProductDescription\" class=\"Block Panel\">\n <h3 class=\"sub-title1\">Gene & Protein 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>UniProt ID</b>\n </div>\n <div class=\"product-tech-info__line-item-right\">\n H2A - P04908 (alt. names: H2A type 1-B/E, H2A.2, H2A/a,\n H2A/m)<br />\n H2B - O60814 (alt. names: H2B K, HIRA-interacting protein 1)<br />\n H3.1 - P68431 (alt. names: H3, H3/a, H3/b, H3/c, H3/d)<br />\n H4 - P62805\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\">References</h3>\n <div\n class=\"ProductDescriptionContainer product-droppdown__section-description\"\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 et al. <em>J. R. Soc. Interface</em> (2013). 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 et al. <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=\"ProductDescriptionContainer product-droppdown__section-description\"\n >\n <div class=\"product-documents\">\n <a\n href=\"/content/documents/tds/16-0029.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=\"16-0029 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, Biotinylated","cart_url":"https://www.epicypher.com/cart.php","max_purchase_quantity":0,"mpn":null,"upc":null,"options":[],"related_products":[{"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 H2AE61A"}],"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":803,"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=864","price":{"without_tax":{"currency":"USD","formatted":"$475.00","value":475},"tax_label":"Sales Tax"},"add_to_wishlist_url":"/wishlist.php?action=add&product_id=864"},{"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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- Species: Human
- Source: E. coli & synthetic DNA
- Tag: Biotinylated
- Molecular Weight: 199,626 Da
Description
Recombinant mononucleosomes consist of 147 base pairs of DNA wrapped around an octamer core of histone proteins (two each of H2A, H2B, H3.1, and H4) to form a nucleosome, the basic repeating unit of chromatin. The 147 bp 601 sequence, which contains a 5’ biotin-TEG group, has high affinity for histone octamers and is useful for nucleosome assembly [1]. Histone H2A contains a glutamate-to-alanine (E-to-A) substitution at position 61 (H2AE61A). H2AE61 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 SMARCA5/SNF2h [4].
Validation Data
Figure 1: Protein gel gata
Coomassie stained PAGE gel of proteins in H2AE61A
mononucleosome (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
and H4) are indicated.
Figure 2: DNA gel data
H2AE61A mononucleosome resolved by native PAGE and stained
with ethidium bromide to visualize DNA.
Lane 1: Free DNA (EpiCypher
18-0005; 100 ng). Lane 2: Intact H2AE61A
mononucleosomes (400 ng).
Figure 3: Functional binding assay
The presence of acidic patch mutations disrupts LANA peptide
binding to recombinant nucleosomes (WT control, EpiCypher
16-0006; H2AE61A, EpiCypher 16-0029; H2AE92K, EpiCypher
16-0030; H2BE105A,E113A, EpiCypher
16-0031). The binding of GST-tagged LANA peptide to biotinylated
recombinant nucleosomes was assessed by AlphaLISA assay
using Streptavidin Donor Beads and Glutathione Acceptor
Beads (PerkinElmer). The presence of H2A acidic patch
mutations completely blocks LANA binding, while H2B
mutations cause a decrease in LANA binding affinity.
Technical Information
Application Notes
H2AE61A mononucleosome is highly purified and suitable for a variety of applications to test the effect of acidic patch mutation on enzymatic activity or chromatin binding. The biotinylated DNA enables affinity binding applications.
Gene & Protein Information
H2B - O60814 (alt. names: H2B K, HIRA-interacting protein 1)
H3.1 - P68431 (alt. names: H3, H3/a, H3/b, H3/c, H3/d)
H4 - P62805