Revision 2

#4083Store at +4C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

IF-IC, FC-FP

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Product Information

Product Usage Information

Directions for Use: DAPI is supplied as a lyophilized powder. For a 20 mg/ml stock, reconstitute the 1 mg in 50 µl deionized water or dimethylformamide. Please follow CST’s recommended IF and Flow protocols. For both applications, following secondary detection:

Immunofluorescence: Counterstain with DAPI as the final step in your staining procedure. Rinse samples twice in PBS for five min each. Dilute DAPI stock solution to a concentration between 1 - 0.1 µg/ml in PBS and incubate for 5 min at room temperature in the dark. Rinse samples once in PBS and then prepare for imaging. Examine immediately using appropriate excitation wavelength.

Alternatively, dilute DAPI stock solution to a concentration between 1 - 0.1 µg/ml in mounting media, apply to cells, and prepare for imaging. Examine immediately using appropriate excitation wavelength.

Flow Cytometry: Rinse samples once in Incubation Buffer. Dilute DAPI stock solution to a concentration between 1.60-0.400 µg/ml in PBS and incubate for 15 min at room temperature in the dark before analyzing cells on flow cytometer.

Storage

Store lyophilized or in solution at 4°C, desiccated. Protect from light. In lyophilized form, the chemical is stable for 24 months. Once in solution, use within 2-3 weeks to prevent loss of potency. For long-term storage, aliquot and store at -20°C to avoid multiple freeze/thaw cycles.

Product Description

DAPI is supplied as a lyophilized powder in 1 mg units. It can be used to examine cellular DNA in fluorescent microscopy and cytometry applications.
Molecular Formula C16H15N5 2(HCl)

Background

4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) is a blue fluorescent DNA dye that targets double-stranded AT clusters in the DNA minor groove (1). One molecule of dye binds to each 3 base pairs of dsDNA and yields an approximate 20-fold fluorescent enhancement (2). The level of DAPI-DNA fluorescence is proportional to DNA content (3).

  1. Portugal, J. and Waring, M.J. (1988) Biochim Biophys Acta 949, 158-68.
  2. Kapuscinski, J. (1995) Biotech Histochem 70, 220-33.
  3. Manzini, G. et al. (1983) Nucleic Acids Res 11, 8861-76.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Applications Key

IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 2
#4083

DAPI

Immunofluorescence Image 1: DAPI Expand Image
使用 COX IV (3E11) Rabbit mAb #4850(绿色)、β-Catenin (L54E2) Mouse mAb (IF Preferred) #2677(红色)和 DAPI(蓝色)对 HeLa 细胞进行免疫荧光分析。
Immunofluorescence Image 2: DAPI Expand Image
使用 VE-Cadherin (D87F2) XP® Rabbit mAb #2500(绿色)和 DAPI(蓝色)对 HUVE 细胞进行免疫荧光分析。肌动蛋白丝用 DyLight 554 Phalloidin #13054(红色)标记。
Flow Cytometry Image 1: DAPI Expand Image
使用 Ki-67 (D3B5) Rabbit mAb (Alexa Fluor® 647 Conjugate) #12075 和 DAPI 对 Jurkat 细胞进行流式细胞分析(DNA 含量)。