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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, Prod # 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 (Prod # A21300) followed by HRP-conjugated goat anti-mouse IgG antibody and Oregon Green® 488 tyramide (TSA Kit #9, Prod # 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 (Prod # A21253) and diaminobenzidine (DAB) staining. Both nuclear and non-nuclear (presumably mitochondrial) incorporation of BrdU is clearly visible in the resulting image.
|Tested species reactivity||Chem|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Storage buffer||0.1M potassium phosphate, pH 8|
|Contains||5mM sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Immunocytochemistry (ICC)||Assay Dependent|
|Immunohistochemistry (IHC)||Assay Dependent|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Anti-fluorescent dye antibodies recognize specific fluorophores and, in most cases, quench their fluorescence. Thus many anti-dye antibodies, including those that recognize fluorescein, can serve as cell-impermeant probes for determining whether fluorescent dye-conjugated ligands, proteins, bacteria or other biomolecules have been internalized by endocytic or pinocytic processes.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
MicroRNA-146a: a key regulator of astrocyte-mediated inflammatory response.
A-21253 was used in immunohistochemistry - paraffin section to assess the role of miR-46a in the modulation of astrocyte-mediated inflammation.
|Iyer A,Zurolo E,Prabowo A,Fluiter K,Spliet WG,van Rijen PC,Gorter JA,Aronica E||PloS one (7:null)||2012|
Multicolor fluorescent in situ hybridization to define abutting and overlapping gene expression in the embryonic zebrafish brain.
A-21253 was used in immunohistochemistry to discuss methods to map overlapping and abutting regulatory gene expression domains.
|Lauter G,Söll I,Hauptmann G||Neural development (6:null)||2011|