Cellular DNA Fragmentation ELISA
ROCHE/11585045001
Synonym: ELISA
Product Type: Chemical
Principle of the Cellular DNA Fragmentation ELISA
Kinetics of camptothecin-induced apoptotic cell death in HL60 cells. 5 x 104 HL60 cells/mL were incubated either in the presence of 200 ng/mL camptothecin (CAM) (red; blue) or absence (yellow; green) for 1 to 8 hours at +37 °C. After the times indicated, 100 μl/well supernatants (SN) (blue; green) were removed and analyzed by the Cellular DNA Fragmentation ELISA. Subsequently, the cells were lysed and the lysates (LYS) (red; yellow) were also tested by the same ELISA.
| manufacturer/tradename | Roche |
| Quality Level | 100  |
| storage temp. | 2-8°C |
| usage | sufficient for ≤500 tests |
| Application: | For research use only. Not for use in diagnostic procedures. The Cellular DNA Fragmentation ELISA is for the quantification of BrdU-labeled DNA fragments either released from cells during necrosis or cell-mediated cytotoxicity, or within the cytoplasm of apoptotic cells. |
| Features and Benefits: | • Nonradioactive assay system • Sensitive (1 x 103 cells/well) • Low assay background • No additional dilution steps required • No washing of the cells required • Results obtained correlate to those obtained by standard methods • Denaturation/fixation of DNA in the microplate by microwave irradiation • Fast (4.5 - 5.5 hours) |
| General description: | The Cellular DNA Fragmentation ELISA is a photometric enzyme-linked immunosorbent assay (ELISA) for the detection of 5’-bromo-2’-deoxy-uridine (BrdU)-labeled DNA fragments in culture supernatants and cell lysates. BrdU is used as a metabolic labeling agent by the nuclear DNA of target cells. This BrdU-labeled DNA can be detected easily and quantified using a monoclonal antibody against BrdU in an ELISA. This kit is a non-radioactive alternative to the [3H]-thymidine release assay, the [51Cr]-release assay, and the [3H]-thymidine DNA fragmentation assay. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Packaging: | 1 kit containing 8 components. |
| Preparation Note: | Working solution: Solubility: soluble in water, aqueous buffers, and dimethylformamide. Can be dissolved in methanol. A clear solution is achieved at 20 mg/ml. Storage conditions (working solution): Storage of coated plates: 1 week at 2 to 8 °C. The anti-BrdU-POD stock solution is stable at 2 to 8 °C for several months; for long- term storage it is recommended to store the solution in aliquots at -15 to -25 °C; The anti-BrdU-working solution should not be stored. For the anti-DNA-clone MCA-33 we do no have stability data. However it should be no problem to aliquot and store this non-conjugated antibody at -15 to -25 °C for longer storage. |
| Symbol |  GHS07 |
| Signal word | Warning |
| Hazard statements | H317 - H319 - H412 |
| Precautionary statements | P261 - P273 - P280 - P333 + P313 - P337 + P313 - P362 + P364 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352200 |
| Components | Anti-DNA Antibody (clone MCA-33); Anti-BrdU Peroxidase antibody, Fab Fragments (clone BMG 6H8); Coating Buffer; Washing Buffer; Incubation Buffer; Substrate Solution; BrdU Labeling Reagent; Adhesive Cover Foils |