Nuclear and nonnuclear incorporation of 5-bromo-2'-deoxyuridine in live cells.

Nuclear and nonnuclear incorporation of 5-bromo-2'-deoxyuridine in live cells. Bovine pulmonary arterial endothelial (BPAE) cells were labeled with 5-bromo-2'-deoxyuridine (BrdU, Cat. no. B23151) applied at a concentration of 10 µM for 30 minutes. After fixation with 4% formaldehyde in phosphate-buffered saline for 30 minutes, chromatin was denatured by treatment with 2 M HCl for 20 minutes. Incorporated BrdU was detected with mouse monoclonal anti-bromodeoxyuridine antibody (Cat. no. A21300) followed by HRP-conjugated goat anti–mouse IgG antibody and Oregon Green® 488 tyramide (TSA Kit #9, Cat. no. T20919). Tyramide labeling was further amplified and converted for visualization by bright-field microscopy by detection of the Oregon Green® 488 dye hapten using the HRP conjugate of anti-fluorescein/Oregon Green® antibody (Cat. no. A21253) and diaminobenzidine (DAB) staining. Both nuclear and non-nuclear (presumably mitochondrial) incorporation of BrdU is clearly visible in the resulting image.

Nuclear and nonnuclear incorporation of 5-bromo-2'-deoxyuridine in live cells. Bovine pulmonary arterial endothelial (BPAE) cells were labeled with 5-bromo-2'-deoxyuridine (BrdU, Cat. no. B23151) applied at a concentration of 10 µM for 30 minutes. After fixation with 4% formaldehyde in phosphate-buffered saline for 30 minutes, chromatin was denatured by treatment with 2 M HCl for 20 minutes. Incorporated BrdU was detected with mouse monoclonal anti-bromodeoxyuridine antibody (Cat. no. A21300) followed by HRP-conjugated goat anti–mouse IgG antibody and Oregon Green® 488 tyramide (TSA Kit #9, Cat. no. T20919). Tyramide labeling was further amplified and converted for visualization by bright-field microscopy by detection of the Oregon Green® 488 dye hapten using the HRP conjugate of anti-fluorescein/Oregon Green® antibody (Cat. no. A21253) and diaminobenzidine (DAB) staining. Both nuclear and non-nuclear (presumably mitochondrial) incorporation of BrdU is clearly visible in the resulting image.

Related Products

Related Images

A prometaphase muntjac skin fibroblast stained with Alexa Fluor® 350 phalloidin, an anti–a-tubulin antibody and an anti–cdc6 peptide antibody. Go ›

Bovine pulmonary artery endothelial cells (BPAEC). MitoTracker® Red CMXRos, SYTOX® Green nucleic acid stain, biotin-XX goat anti–mouse IgG antibody and Cascade Blue® NeutrAvidin biotin-binding protein. Go ›

BPAE cells fixed and permeabilized using the Image-iT® Fixation/Permeabilization Kit. Go ›

BPAE cells fixed and permeabilized using the Image-iT® Fixation/Permeabilization Kit. Go ›

Agarose gel containing camptothecin-treated HL-60 cells. Go ›

Imaging autophagy in live HeLa cells with CellLight® reagents for mitochondria and lysosomes: Go ›

Simultaneous detection of expression of five genes in a whole mount Drosophila embryo by fluorescence in situ hybridization (FISH) with five RNA probes Go ›

Immunocytochemistry using Alpha-Tubulin Monoclonal Antibody, Mouse Go ›