|Related Categories||Apoptosis Detection, Apoptosis and Cell Cycle, Cell Analysis, Cell Biology, Cell Signaling and Neuroscience,|
|usage||sufficient for 60 cell suspensions|
|shipped in||wet ice|
The greater incorporation of Br-dUTP results in a stronger signal by flow cytometry when detected using a fluorescein-labeled anti-BrdU antibody. Propidium iodide/RNase A solution is included in the kit to counterstain the total DNA.
Greater incorporation of Br-dUTP, resulting in improved detection than found by using biotin- or digoxigenin-conjugated dUTP or by using fluorochrome (fluorescein or BODIPY)-conjugated deoxynucleotides.
The kit is shipped in two parts, both wet ice. Upon receipt, store APO-PART1 at −20 °C and APO-PART2 at 2-4 °C
Br-dUTP is incorporated more readily into the DNA fragments than deoxyuridine triphosphate labeled with larger dyes such as FITC, biotin or digoxigenin. One of the biological characteristics that defines apoptosis is the degradation of genomic DNA into fragments of 180-200 bp, commonly called "DNA laddering". The fragmentation creates a large number of 3′-hydroxyl sites at the DNA breaks. This property is used in the APO-BRDU kit to identify apoptotic cells by labeling the 3′-hydroxyl sites with bromodeoxyuridine triphosphate (Br-dUTP). Br-dUTP is enzymatically attached to the 3-hydroxyl sites of double- or single-stranded DNA by terminal transferase (TdT). Non-apoptotic cells do not incorporate Br-dUTP due to the lack of available 3-hydroxyl sites.
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|Br-dUTP 480 μL|
|Fluorescein PRB-1 antibody 300 μL|
|Negative control cells 5 mL|
|PI/RNase staining buffer 30 mL|
|Positive control cells 5 mL|
|Reaction buffer 0.6 mL|
|Rinsing buffer 120 mL|
|Terminal deoxynucleotidyl transferase (TdT) 45 μL|
|Wash buffer 120 mL|
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