{"product_id":"dapi-reagent-25-g-ml","title":"DAPI Reagent (25μg\/mL)","description":"\u003ch2\u003eDAPI Reagent (25μg\/mL)\u003c\/h2\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eIntroduction\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003e\u003cspan\u003eElabscience\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e®\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003e  DAPI Reagent (25μg\/mL) is developed to identify apoptotic and necrotic cells.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp align=\"left\" class=\"MsoNormal\"\u003e\u003cspan\u003eDAPI Reagent has a high DNA binding constant and is efficiently excluded by intact cells. It is useful for DNA analysis and dead cell discrimination during flow cytometric analysis. \u003c\/span\u003e\u003cspan\u003eDue to the loss of integrity of membrane, DAPI can enter late apoptotic or necrotic cells to stain DNA. Cells at different apoptotic stages can be distinguished by using \u003c\/span\u003e\u003cspan\u003eDAPI\u003c\/span\u003e\u003cspan\u003e and Annexin V.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003e\u003ci\u003e\u003cspan\u003eJurkat cells were treated with 5 μM Camptothecin and detected with this reagent and Annexin V-APC.\u003c\/span\u003e\u003c\/i\u003e\u003c\/p\u003e\n\u003cp class=\"MsoNormal\"\u003e\u003cspan\u003eJurkat cells were cultured with (\u003c\/span\u003e\u003cb\u003e\u003cspan\u003eRight\u003c\/span\u003e\u003c\/b\u003e\u003cspan\u003e) or without (\u003c\/span\u003e\u003cb\u003e\u003cspan\u003eLeft\u003c\/span\u003e\u003c\/b\u003e\u003cspan\u003e)  5 μM Camptothecin for 4 h. Annexin V-APC single-positive cells were early apoptotic cells, Annexin V-APC and DAPI double-positive cells were necrotic or late apoptotic cells, and DAPI single-positive cells were naked nuclei.\u003c\/span\u003e\u003c\/p\u003e\n\u003cdiv class=\"detail_section\"\u003e\u003cbr\u003e\u003c\/div\u003e\n\u003cp\u003e\u003cstrong\u003eStaining Procedure\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\u003c\/ol\u003e\n\u003col\u003e\n\u003cli\u003e\n\u003cspan\u003eInduce apoptosis of suspension cells with reagents of interest. Collect cell cultures, centrifuge at 300 g for 5 min and discard the supernatant. Add PBS to wash the cells and resuspend the cells gently followed by the cell counting.\u003cbr\u003e\u003c\/span\u003eTip: This product is only validated in suspension cells. Good cell viability is the key to the experiment. When the adherent cells are used for apoptotic detection, treatments like digestion may increase the ratio of necrotic or apoptotic cells and cause uncontrollable effects on the experimental results. Please be aware！\u003c\/li\u003e\n\u003cli\u003e\n\u003cspan\u003eSplit the cell suspension into tubes, 1~5 × 10\u003c\/span\u003e\u003csup\u003e\u003cspan\u003e5\u003c\/span\u003e\u003c\/sup\u003e\u003cspan\u003e\u003csup\u003e \u003c\/sup\u003ecells for each, centrifuge at 300 g for 5 min and discard the supernatant. Add PBS to wash the cells and discard the supernatant. Add 500 μL of 1 × Annexin V Binding Buffer[\u003ca href=\"https:\/\/www.elabscience.com\/p-annexin_v_binding_buffer(10%C3%97)[151]-205765.html\" target=\"_blank\"\u003eE-CK-A151\u003c\/a\u003e] to resuspend the cells.\u003c\/span\u003e\n\u003c\/li\u003e\n\u003cli\u003e\u003cspan\u003eAdd 5 µl of Annexin V-APC[\u003ca href=\"https:\/\/www.elabscience.com\/p-annexin_v_apc_reagent[117]-205757.html\" target=\"_blank\"\u003eE-CK-A117\u003c\/a\u003e] and 5 µl of  DAPI Reagent (25μg\/mL) to each tube.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan\u003eGently vortex the cells and incubate at room temperature for 15~20 min in the dark.\u003c\/span\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cspan\u003eAnalyze the cells immediately with proper machine settings. Otherwise, place the cells on ice in the dark and analyze within 1 h.\u003c\/span\u003e\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eStorage\u003c\/strong\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan\u003eStore at \u003c\/span\u003e\u003cspan\u003e2~8°C\u003c\/span\u003e\u003cspan\u003e for one year in dark.  DAPI Reagent (25μg\/mL) should be spilt into small tubes.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eNotes\u003c\/strong\u003e\u003c\/p\u003e\n\u003col\u003e\n\u003cli\u003e\n\u003cp style=\"font-size: 0.875rem; display: inline !important;\" class=\"MsoListParagraph\"\u003eFor maximal assay performance, this reagent should be used within 12 months. Avoid freeze \/ thaw cycles.\u003c\/p\u003e\n\u003c\/li\u003e\n\u003cli\u003eDetect apoptosis as soon as possible after staining to avoid the increase number of apoptosis or necrosis.\u003c\/li\u003e\n\u003cli\u003eAvoid extended exposure of the samples to direct light to protect the fluorophores from quenching.\u003c\/li\u003e\n\u003cli\u003eFor your safety and health, please wear the lab coat and disposable gloves before the experiments.\u003c\/li\u003e\n\u003c\/ol\u003e","brand":"Elabscience Biotechnology","offers":[{"title":"500 Tests","offer_id":40441418448954,"sku":"E-CK-A163-500","price":127.95,"currency_code":"USD","in_stock":true},{"title":"200 Tests","offer_id":40441418481722,"sku":"E-CK-A163-200","price":94.95,"currency_code":"USD","in_stock":true},{"title":"100 Tests","offer_id":40441418514490,"sku":"E-CK-A163-100","price":83.95,"currency_code":"USD","in_stock":true},{"title":"50 Tests","offer_id":40441418547258,"sku":"E-CK-A163-50","price":77.95,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0722\/7785\/files\/E-CK-A163.jpg?v=1714097074","url":"https:\/\/www.msesupplies.com\/en-de\/products\/dapi-reagent-25-g-ml","provider":"MSE Supplies","version":"1.0","type":"link"}