Exponentially growing promyelocytic leukemia (HL60) cells. Hoechst 33258 and ChromaTide BODIPY FL-14-dUTP.

Detection of BrdU-incorporating cells using the SBIP (Strand Breaks Induced by Photolysis) technique. Exponentially growing human promyelocytic leukemia (HL60) cells were incubated with 20 µM BrdU (B23151) for 40 minutes and then with 20 µg/mL Hoechst 33258 (H1398, H3569, H21491) in the presence of 2% DMSO for an additional 20 minutes. After this incubation, the cells were exposed to 300 nm UV light for 5 minutes to selectively photolyze DNA that contained the incorporated BrdU, and then fixed in 70% ethanol. Subsequent incubation of the permeabilized cells with the ChromaTide BODIPY FL-14-dUTP (C7614) in the presence of exogenous terminal deoxynucleotidyl transferase resulted in incorporation of the fluorophore into DNA strand breaks, thereby labeling the S-phase cells. The DNA was counterstained with the red-fluorescent nucleic acid stain propidium iodide (P1304MP, P3566, P21493); therefore, the BODIPY FL dye–labeled DNA appears yellow. The image was contributed by Zbigniew Darzynkiewicz, Cancer Research Institute, New York Medical College.

Detection of BrdU-incorporating cells using the SBIP (Strand Breaks Induced by Photolysis) technique. Exponentially growing human promyelocytic leukemia (HL60) cells were incubated with 20 µM BrdU (B23151) for 40 minutes and then with 20 µg/mL Hoechst 33258 (H1398, H3569, H21491) in the presence of 2% DMSO for an additional 20 minutes. After this incubation, the cells were exposed to 300 nm UV light for 5 minutes to selectively photolyze DNA that contained the incorporated BrdU, and then fixed in 70% ethanol. Subsequent incubation of the permeabilized cells with the ChromaTide BODIPY FL-14-dUTP (C7614) in the presence of exogenous terminal deoxynucleotidyl transferase resulted in incorporation of the fluorophore into DNA strand breaks, thereby labeling the S-phase cells. The DNA was counterstained with the red-fluorescent nucleic acid stain propidium iodide (P1304MP, P3566, P21493); therefore, the BODIPY FL dye–labeled DNA appears yellow. The image was contributed by Zbigniew Darzynkiewicz, Cancer Research Institute, New York Medical College.

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Sku Name Size Price Qty
B23151 BrdU (5-Bromo-2´-Deoxyuridine) 100 mg USD 96.50
H1398 Hoechst 33258, Pentahydrate (bis-Benzimide), 100 mg 100 mg USD 63.25
H21491 Hoechst 33258, Pentahydrate (bis-Benzimide) - FluoroPure™ Grade 100 mg USD 74.00
H3569 Hoechst 33258, Pentahydrate (bis-Benzimide) - 10 mg⁄mL Solution in Water 10 mL USD 84.00
P1304MP Propidium Iodide 100 mg USD 129.00
P21493 Propidium Iodide - FluoroPure™ Grade 100 mg USD 154.00
P3566 Propidium Iodide - 1.0 mg⁄mL Solution in Water 10 mL USD 68.25

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