DAPI Permeability 2020

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December DAPI Permeability 2020 DAPI (4′,6-diamidino-2-phenylindole) is a fluorescent stain often used to differentiate between live and dead cells for viability measurements in flow cytometry. This reagent is a popular choice due to its short incubation time and high relative brightness. Similar to other reagents, DAPI staining conditions must be optimized for best experimental results. Incubation Conditions Concentration Live cell Live cell DAPI MFI DAPI MFI 1 hour on ice 582 582 1.0 Time 0 Room Temp 629 0.2 241 1 hour (µg/mL) ConcentrationDAPI Incubation Time Temperature & Incubation Room Temp 2,480 0.1 259 DAPI MFI DAPI MFI Samples in figure above were all stained at a final concentration of 1.0 µg/mL. Figure above shows the the impact of staining with increasing concentrations of When left at room temperature for 1 h the live population experienced a shift in DAPI. All samples were kept on ice and acquired immediately after DAPI was DAPI signal, whereas the sample kept on ice did not, indicating colder added. 1.0 µg/mL shows a shift of the live cell population, indicating the temperatures can be beneficial in preventing DAPI uptake in viable cells. concentration is too high. DAPI is a fast acting, bright fluorochrome which binds to the Adenine-Thymine regions in dsDNA. It is important to remember that DAPI can be permeable to live cells. DAPI uptake by live cells can be prevented by optimizing concentration, adding it immediately prior to acquisition and controlling sample temperature, understanding that ideal conditions can vary across cell types. The FCCF thanks Sara Haddock for providing fcs files used in this Post-it https://fccf.mskcc.org.

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DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) for Nucleic Acid Staining
DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) for nucleic acid staining Catalog number: AR1176-10 Boster’s DAPI solution is a fluorescent dye with higher efficiency for immunofluorescence microscopy. This package insert must be read in its entirety before using this product. For research use only. Not for use in diagnostic procedures. BOSTER BIOLOGICAL TECHNOLOGY 3942 B Valley Ave, Pleasanton, CA 94566 Phone: 888-466-3604 Fax: 925-215-2184 Email:[email protected] Web: www.bosterbio.com DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) for nucleic acid staining Catalog Number: AR1176-10 Product Overview Material DAPI dihydrochloride (MW = 350.3) Form Liquid Size 10 mL(100 assys) Concentration 1μg/ml 8 mM sodium phosphate, 2 mM potassium phosphate, 140 mM sodium chloride, 10 Buffer mM potassium chloride; pH 7.4. Storage Upon receipt store at -20°C, protect from light. Stability When stored as directed, product should be stable for one year. Molecular formula C16H15N5 • 2 HCL Excitation: DAPI 340nm Emission: DAPI 488nm Excitation: DAPI-DNA complexes 360nm Emission: DAPI-DNA complexes 460nm Thermofisher (Product No. 62247); Thermofisher (Product No. 62248); Millipore Equivalent Sigma (Product No. D9542) BOSTER BIOLOGICAL TECHNOLOGY 3942 B Valley Ave, Pleasanton, CA 94566 Phone: 888-466-3604 Fax: 925-215-2184 Email:[email protected] Web: www.bosterbio.com Notes: Type of DAPI Molecular formula Molecular weight Catalog Number DAPI dihydrochloride C16H15N5 • 2 HCL 350.25 AR1176-10 DAPI dilactate C16H15N5 • 2 C3H6O3 457.48 N/A Introduction DAPI (4',6-diamidino-2-phenylindole) is a fluorescent dye which can bind DNA strands robustly, the fluorescence being detected by fluorescence microscope.
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