647 EdU Click Proliferation

The BD Pharmingen™ 647 EdU Click Proliferation Kit provides a tool for analyzing DNA synthesis in cycling cells. In this method, the thymidine analog EdU (5-ethynyl-2´-deoxyuridine) is incorporated into newly synthesized DNA by cells progressing through the S (DNA synthesis) phase of the cell cycle. EdU can then be detected in fixed and permeabilized cells with a fluorophore-labeled azide. The reaction of the fluorophore-labeled azide and EdU occurs via a copper-catalyzed click reaction between the azide moiety of the fluorophore and the alkyne moiety of EdU. When co-stained with a DNA dye such as 7-AAD, PI, or DAPI, cell populations may be segmented by flow cytometry into the G0/G1-phases (2N DNA content, EdU-negative), S-phase (2N-4N DNA content, EdU-positive), or G2/M-phases (4N DNA content, EdU-negative). The BD Pharmingen™ 647 EdU Click Proliferation Kit contains  Eterneon™ Red 645 Azide, which is excited by the red laser and has an excitation maximum of 643 nm and an emission maximum of 662 nm. Please note that this kit is provided as Part 1 of 2 (Components A, B, and C) to be stored dry and protected from light at -20°C, and Part 2 of 2 (Components D, E, F, and G) to be stored dry at 2 - 8°C or room temperature. See "Kit Contents" immediately below for more information. Kit Contents Component ID          Component Description          Amount          Long Term Storage Conditions     A          EdU (5-ethynyl-2´-deoxyuridine)            10 mg               -20°C, dry     B          Eterneon™ Red 645 azide (10 mM)          130 μL               -20°C, dry, dark     C          Buffer Additive (10×)          400 mg               -20°C, dry     D          Saponin-based Permeabilization and Wash Reagent (10×)            50 mL                  2 - 8°C     E          Fixative Solution (4% paraformaldehyde-based)              5 mL                  2 - 8°C     F          Catalyst Solution              2 mL                 RT, dry     G          DMSO              5 mL                 RT, dry Please note that Hazard Warnings for components listed above are found on Page 5 of this document.

The BD Pharmingen™ 647 EdU Click Proliferation Kit provides a tool for analyzing DNA synthesis in cycling cells. In this method, the thymidine analog EdU (5-ethynyl-2´-deoxyuridine) is incorporated into newly synthesized DNA by cells progressing through the S (DNA synthesis) phase of the cell cycle. EdU can then be detected in fixed and permeabilized cells with a fluorophore-labeled azide. The reaction of the fluorophore-labeled azide and EdU occurs via a copper-catalyzed click reaction between the azide moiety of the fluorophore and the alkyne moiety of EdU. When co-stained with a DNA dye such as 7-AAD, PI, or DAPI, cell populations may be segmented by flow cytometry into the G0/G1-phases (2N DNA content, EdU-negative), S-phase (2N-4N DNA content, EdU-positive), or G2/M-phases (4N DNA content, EdU-negative). The BD Pharmingen™ 647 EdU Click Proliferation Kit contains  Eterneon™ Red 645 Azide, which is excited by the red laser and has an excitation maximum of 643 nm and an emission maximum of 662 nm. Please note that this kit is provided as Part 1 of 2 (Components A, B, and C) to be stored dry and protected from light at -20°C, and Part 2 of 2 (Components D, E, F, and G) to be stored dry at 2 - 8°C or room temperature. See "Kit Contents" immediately below for more information. Kit Contents Component ID          Component Description          Amount          Long Term Storage Conditions     A          EdU (5-ethynyl-2´-deoxyuridine)            10 mg               -20°C, dry     B          Eterneon™ Red 645 azide (10 mM)          130 μL               -20°C, dry, dark     C          Buffer Additive (10×)          400 mg               -20°C, dry     D          Saponin-based Permeabilization and Wash Reagent (10×)            50 mL                  2 - 8°C     E          Fixative Solution (4% paraformaldehyde-based)              5 mL                  2 - 8°C     F          Catalyst Solution              2 mL                 RT, dry     G          DMSO              5 mL                 RT, dry Please note that Hazard Warnings for components listed above are found on Page 5 of this document.

51-9011284AK/(2 to 8C)+51-9011285BK/(max.-20C)

Intracellular staining (flow cytometry), Immunocytochemistry, Immunofluorescence (Tested During Development)

研发参考(6) 1. Buck SD, Bradford J, Gee KR, Agnew BJ, Clarke ST, Salic A. Detection of S-phase cell cycle progression using 5-ethynyl-2’-deoxyuridine incorporation with click chemistry, an alternative to using 5-bromo-2’-deoxyuridine antibodies. Biotechniques. 2008; 44(7):927-929. (Methodology). 2. Cavanagh BL, Walker T, Norazit A, Meedeniya AC. Thymidine analogues for tracking DNA synthesis. Molecules. 2011; 16(9):7980-7993. (Methodology). 3. Kolb HC, Finn MG, Sharpless KB. Click Chemistry: Diverse Chemical Function from a Few Good Reactions. Angew Chem Int Ed Engl. 2001; 40(11):2004-2021. (Methodology). 4. Kotogany E, Dudits D, Horvath GV, Ayaydin F. A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine. Plant Methods. 6(1)(Methodology). 5. Salic A, Mitchison TJ. A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc Natl Acad Sci U S A. 2008; 105(7):2415-2420. (Methodology). 6. Wang Q, Chan TR, Hilgraf R, Fokin VV, Sharpless KB, Finn MG. Bioconjugation by Copper(I)-Catalyzed Azide-Alkyne [3 + 2] Cycloaddition. J Am Chem Soc. 2003; 125(11):3192-3193. (Methodology).