APOBRDU

Sigma-Aldrich

Flow Cytometry Kit for Apoptosis

NACRES:
NA.32
Pricing and availability is not currently available.

usage

sufficient for 60 cell suspensions

packaging

pkg of 1 kit

application(s)

cell analysis: suitable
detection: suitable
flow cytometry: suitable

detection method

fluorometric

shipped in

wet ice

storage temp.

2-8°C

Related Categories

Application

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. APOBRDU has been used in flow cytometry assay in human non–small cell lung cancer. APOBRDU has been used in cell cycle and apoptosis assay in HeLa cells, and in tunel assay human neuroblastoma cell lines.

Features and Benefits

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.

Packaging

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

Principle

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.

Kit Components Only

Product No.
Description

  • 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 .6 mL

  • Rinsing buffer 120 mL

  • Terminal deoxynucleotidyl transferase (TdT) 45 μL

  • Wash buffer 120 mL

See All (9)

Signal Word

Danger

Hazard Statements

Hazard Codes

F,T

Risk Statement

11-23/25

Safety Statement

16-45

RIDADR

UN 3316 9

WGK Germany

WGK 3

Flash Point(F)

55.4 °F

Flash Point(C)

13 °C

Certificate of Analysis
Certificate of Origin
X Li et al.
Cell proliferation, 28(11), 571-579 (1995-11-01)
In situ presence of numerous DNA strand breaks is a typical feature of apoptotic cells. Selective DNA strand break induction by photolysis (SBIP) at sites that contain incorporated halogenated DNA precursors has recently been proposed as a method of analysing...
Cardenolide-induced lysosomal membrane permeabilization demonstrates therapeutic benefits in experimental human non-small cell lung cancers
Mijatovic T, et al.
Neoplasia, 8(5), 402-412 (2006)
The novel lipophilic camptothecin analogue gimatecan is very active in vitro in human neuroblastoma: a comparative study with SN38 and topotecan
Di Francesco AM, et al.
Biochemical Pharmacology, 70(8), 1125-1136 (2005)
Cdc7 is an active kinase in human cancer cells undergoing replication stress
Tenca P, et al.
The Journal of Biological Chemistry, 282(1), 208-215 (2007)
Articles
Cell based assays for cell proliferation (BrdU, MTT, WST1), cell viability and cytotoxicity experiments for applications in cancer, neuroscience and stem cell research.
Read More

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