{"product_id":"nuclear-extract-kit","title":"Nuclear Extract Kit","description":"\u003cdiv class=\"fb_smallest fb_8-12_smallest margin-bottom-normal\"\u003e\n\u003cdiv class=\"margin-bottom-normal\"\u003e\n\u003cp\u003eThe Nuclear Extract Kit isolates high-quality nuclear, cytoplasmic or whole-cell extract from mammalian cell or tissue samples. This eliminates the need for optimizing reagents and protocols, and ensures high yields every time. More importantly, standardizing your extraction procedure with our high-quality reagents ensures more reproducible results. Extracts are suitable for use in a various applications including EMSA, Western blot and binding assays and is ideally suited for preparation of extracts for our\u003cspan\u003e \u003c\/span\u003e\u003ca href=\"https:\/\/www.activemotif.com\/catalog\/180\/transam-transcription-factor-elisas\" title=\"Click here to learn more about TransAM Transcription Factor Assays\"\u003eTransAM\u003csup\u003e®\u003c\/sup\u003e\u003cspan\u003e \u003c\/span\u003eTranscription Factor Assays\u003c\/a\u003e.\u003c\/p\u003e\n\u003cp\u003eThe Nuclear Extract Kit works with fresh or frozen cell and tissue samples and yields protein amounts of approximately 0.5 to 1 mg from cytoplasmic extract (at 1-2 mg\/ml concentration), 150 to 250 μg from nuclear extract (at 3-5 mg\/ml concentration), 1.2 to 2.4 mg from whole cell extract (at 4-8 mg\/ml concentration) from 8.8 x 10\u003csup\u003e6\u003c\/sup\u003e\u003cspan\u003e \u003c\/span\u003ecells. The kit procedure is simple and only takes two hours to complete. The kit includes buffers, detergents and reagents for lysis as well as Protease and Phosphatase inhibitors to ensure preservation of your proteins.\u003c\/p\u003e\n\u003ch3\u003eApplications\u003c\/h3\u003e\n\u003cp\u003eExtracts prepared using the Nuclear Extract Kit can be used with Active Motif’s\u003cspan\u003e \u003c\/span\u003e\u003ca href=\"https:\/\/www.activemotif.com\/catalog\/180\/transam-transcription-factor-elisas\" title=\"TransAM® Kits\"\u003eTransAM\u003csup\u003e®\u003c\/sup\u003e\u003cspan\u003e \u003c\/span\u003eKits\u003c\/a\u003e, or in other procedures that require cell extracts, like gelshift assays, Western blots, DNA footprinting, as a starting point for protein purification for binding or activity assays or or other analysis techniques.\u003c\/p\u003e\n\u003cdiv class=\"aname-nav\"\u003e\n\u003ch3\u003eNuclear Extract Kit Advantages:\u003c\/h3\u003e\n\u003cul\u003e\n\u003cli\u003eQuality-controlled reagents ensure reproducibility\u003c\/li\u003e\n\u003cli\u003eNo need to optimize your own procedure\u003c\/li\u003e\n\u003cli\u003eComplete kit contains all required reagents\u003c\/li\u003e\n\u003cli\u003ePrepare nuclear, cytoplasmic or whole-cell extracts with one kit\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"center\"\u003e\n\u003ca href=\"https:\/\/www.activemotif.com\/images\/products\/WB-data_NEK_web.jpg\" target=\"new\"\u003e\u003cimg src=\"https:\/\/www.activemotif.com\/images\/products\/WB-data_NEK_web.jpg\" data-src=\"\/images\/products\/WB-data_NEK_web.jpg\" alt=\"NEK Fractionation Western blots\"\u003e\u003c\/a\u003e\n\u003cp\u003e\u003cspan class=\"caption\"\u003e\u003cb\u003eNEK Fractionation Western blots\u003cbr\u003e(Click image to enlarge)\u003c\/b\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"anchor\" id=\"contents\"\u003e\u003cbr\u003e\u003c\/div\u003e\n\u003cspan\u003e\u003c\/span\u003e\n\u003ch2 class=\"section-title\"\u003eNuclear Extract Kit Contents\u003c\/h2\u003e\n\u003ch3\u003eContents \u0026amp; Storage\u003c\/h3\u003e\n\u003cp\u003eThe Nuclear Extraction Kit is shipped on dry ice and contains reagents with multiple storage temperatures inside. Please store each component at the temperature indicated below. All reagents are guaranteed stable for 6 months from date of receipt when stored properly.\u003c\/p\u003e\n\u003cul\u003e\n\u003cli\u003e1 M DTT (Dithiothreitol); Store at -20°C\u003c\/li\u003e\n\u003cli\u003eProtease Inhibitor Cocktail (PIC); Store at -20°C\u003c\/li\u003e\n\u003cli\u003eLysis Buffer AM1; Store at 4°C\u003c\/li\u003e\n\u003cli\u003e10X PBS; Store at 4°C\u003c\/li\u003e\n\u003cli\u003ePhosphatase Inhibitors; Store at 4°C\u003c\/li\u003e\n\u003cli\u003e10X Hypotonic Buffer; Store at 4°C\u003c\/li\u003e\n\u003cli\u003eDetergent; Store at 4°C\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cdiv class=\"anchor\" id=\"data\"\u003e\u003cbr\u003e\u003c\/div\u003e\n\u003cspan\u003e\u003c\/span\u003e\n\u003ch2 class=\"section-title\"\u003eNuclear Extract Kit Data\u003c\/h2\u003e\n\u003cp\u003eTo demonstrate the efficiency of cellular fractionation using the Nuclear Extract Kit, nuclear, cytoplasmic, and whole-cell extracts were prepared from HeLa cell samples. Nuclear pellets from the nuclear extraction were also collected. Cellular fractions were analyzed by Western blot using antibodies against various proteins. As shown in Figure 1, proteins that serve as markers for cellular fractionation, such as nuclear marker RNA pol II and cytoplasmic marker alpha Tubulin, were only detected in their respective cellular fractions and in whole-cell extracts. The localization of other proteins, such as membrane-associated Lamin A\/C and pan-Cadherin, mitochondrial COX IV and GAPDH were also analyzed.\u003c\/p\u003e\n\u003cdiv class=\"fb_mobile fb_6-12_mobile fb_6-12_tablet fb_4-12_desktop margin-bottom-normal\"\u003e\n\u003cdiv class=\"center margin-bottom-normal\"\u003e\u003cimg src=\"https:\/\/www.activemotif.com\/images\/products\/WB-data_NEK_web.jpg\" height=\"400\" width=\"318\" data-src=\"\/images\/products\/WB-data_NEK_web.jpg\" alt=\"Western blots of specific proteins from cellular fractions obtained with the Nuclear Extract Kit.\"\u003e\u003c\/div\u003e\n\u003cdiv\u003e\n\u003cp\u003e\u003cspan class=\"caption\"\u003e\u003cstrong\u003eFigure 1: Western blots of specific proteins from cellular fractions obtained with the Nuclear Extract Kit.\u003c\/strong\u003e\u003cbr\u003eTo evaluate the efficiency of cellular fractionation, 20 μg of nuclear (Nu), cytoplasmic (Cy) and whole-cell (WC) extracts, along with nuclear pellet collected during nuclear extraction using the Nuclear Extract Kit were analyzed by Western blot using antibodies against various specific proteins. Primary antibodies: Active Motif alpha Tubulin, Catalog No. 39528; COXIV; GAPDH; Active Motif Lamin A\/C, Catalog No. 39288; Pan-Cadherin; Active Motif RNA pol II, Catalog No. 39097.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003chr\u003e\n\u003cp\u003eTo demonstrate the quality of the materials and protocol of the Nuclear Extract Kit, nuclear and cytoplasmic extracts were prepared from HeLa cell samples that were either unstimulated or stimulated with TNF-α for 30 minutes. Because activated NFκB translocates into the nucleus, only nuclear extract from stimulated cells should contain activated NFκB. As predicted, this is precisely what was seen (Figure 2), demonstrating the kit’s high specificity.\u003c\/p\u003e\n\u003cdiv class=\"fb_mobile fb_6-12_mobile fb_6-12_tablet fb_6-12_desktop margin-bottom-normal\"\u003e\n\u003cdiv class=\"center margin-bottom-normal\"\u003e\u003cimg src=\"https:\/\/www.activemotif.com\/images\/products\/nuclear-cyto_compare_v2.jpg\" height=\"221\" width=\"350\" data-src=\"\/images\/products\/nuclear-cyto_compare_v2.jpg\" alt=\"Specific extraction of nuclear and cytoplasmic extracts using the Nuclear Extract Kit\"\u003e\u003c\/div\u003e\n\u003cdiv\u003e\n\u003cp\u003e\u003cspan class=\"caption\"\u003e\u003cstrong\u003eFigure 2: Specific extraction of nuclear and cytoplasmic extracts.\u003c\/strong\u003e\u003cbr\u003eNFκB activation was assayed with the\u003cspan\u003e \u003c\/span\u003e\u003ca href=\"https:\/\/www.activemotif.com\/catalog\/216\/transam-nf%CE%BAb-p50-p52-p65-family-kits\" title=\"TransAM® NFκB p50 Kit\"\u003eTransAM\u003csup\u003e®\u003c\/sup\u003e\u003cspan\u003e \u003c\/span\u003eNFκB p50 Kit\u003c\/a\u003e\u003cspan\u003e \u003c\/span\u003eusing increasing amounts of nuclear and cytoplasmic extracts isolated from unstimulated and stimulated HeLa cells using the Nuclear Extract Kit.\u003c\/span\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cp\u003eActive Motif also offers a number of\u003cspan\u003e \u003c\/span\u003e\u003ca href=\"https:\/\/www.activemotif.com\/catalog\/65\/cell-extracts\" title=\"ready-to-use nuclear, cytoplasmic and whole-cell extracts\"\u003eready-to-use nuclear, cytoplasmic and whole-cell extracts\u003c\/a\u003e. These extracts have been prepared using the same high-quality components found in the Nuclear Extract, and many are supplied as positive controls in TransAM Transcription Factor DNA-binding ELISAs.\u003c\/p\u003e","brand":"ActiveMotif","offers":[{"title":"Default Title","offer_id":44918779674848,"sku":"40410","price":0.0,"currency_code":"KRW","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0601\/9330\/8896\/files\/40010-Nuclear_20Extract_20Kit-0480.jpg?v=1709845012","url":"https:\/\/rndmate.com\/products\/nuclear-extract-kit","provider":"알앤디메이트","version":"1.0","type":"link"}