Cell Death Detection ELISAPLUS
ROCHE/CELLDETH-RO - sufficient for 96 multiwell tests (11774425001), sufficient for 10 x 96 multiwell tests (11920685001)
Synonym: ELISA
Product Type: Chemical
Cell Death Detection ELISAPLUS
Schematic showing the principle of the Cell Death Detection ELISAPLUS.
Dose-dependent induction of apoptosis in U937 cells, detected using the Cell Death Detection ELISAPLUS U937 cells (104 cells/well, in 200 μL) were incubated with different concentrations of camptothecin (CAM) for 4 hours at +37 °C. Before and after lysis, cells were centrifuged and the supernatant was analyzed. Results were plotted as dose vs. response. Result: Amounts of cytoplasmic oligonucleosomes (an indicator of apoptosis) increase as CAM concentration increases. Cell culture supernatants removed from the cells after treatment (but before lysis) gave no signal, indicating that there were no necrotic cells during the treatment.
| manufacturer/tradename | Roche |
| Quality Level | 100  |
| storage temp. | 2-8°C |
| usage | sufficient for 10 x 96 multiwell tests (11920685001) |
| sufficient for 96 multiwell tests (11774425001) |
| Application: | For research use only. Not for use in diagnostic procedures. The Cell Death Detection ELISAPLUS photometric enzyme immunoassay is used for the quantitative in vitro determination of cytoplasmic histone-associated DNA fragments (mono- and oligonucleosomes) after induced cell death. The kit contains a stop solution which allows users to terminate the substrate reaction and run the assay under defined conditions, making it suitable for use in high-throughput applications. |
| Features and Benefits: | • One-step ELISA • High sensitivity: (5 x 102 cells/ml) • Fast performance: (3 - 4 hours) • Positive control included • No prelabeling of cells necessary • Nonradioactive assay system • No species restriction • Easy handling • Low background • Suppressed human anti-mouse factor • Function tested |
| General description: | Use this kit for relative quantification of histone-complexed DNA fragments (mono- and oligonucleosomes) out of the cytoplasma of cells after the induction of apoptosis or when released from necrotic cells. Since the assay does not require prelabeling of cells, it can detect internucleosomal degradation of genomic DNA during apoptosis even in cells that do not proliferate in vitro, for example, freshly isolated tumor cells. The antibodies used in the assay are not species-specific, therefore, the kit may be used to assay cells from a wide variety of species. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Packaging: | • 11774425001 (1 kit containing 10 components) • 11920685001 (1 kit containing 10 components) |
| Packaging: | 1 kit containing 10 components |
| Preparation Note: | Working solution: Preparation of Working Solutions Alongside the ready-to-use solutions supplied with this kit, you will need to prepare the following working solutions: Note: Use only double-distilled water for reconstitution of lyophilizates. Anti-Histone Biotin Reconstitute the lyophilizate in 450 μl double-distilled water for 10 minutes and mix thoroughly. Use: Component of the Immunoreagent. Anti-DNA Peroxidase Reconstitute the lyophilizate in 450 μl double-distilled water for 10 minutes and mix thoroughly. Use: Component of the Immunoreagent. Positive Control Reconstitute the lyophilizate in 450 μl double-distilled water for 10 minutes and mix thoroughly. Use: ELISA Step 1. ABTS Tablets Dependent on the number of samples tested, dissolve 1, 2, or 3 tablets from bottle 7 in 5, 10, or 15 ml Substrate Buffer. Use: ELISA Step 5. ABTS Stop Solution If turbidity or a precipitate is visible, warm to 37 °C with shaking until the solution is clear. Use: ELISA Step 6. Storage conditions (working solution): Anti-Histone Biotin: 2 to 8 °C for 2 months ABTS Tablets: 1 month stored protected from light. Allow to come to 15 to 25 °C before use. ABTS Stop Solution: 2 to 8 °C until the expiration date printed on the label. |
| Preparation Note: | The exact detection limit of dying/dead cells in a particular sample strongly depends on the kinetics of cell death, the cytotoxic agent used, and the amount of affected cells in the total cell population. Using U937/camptothecin (CAM) as a cellular model system for cell death, the immunoassay allows the specific detection of mono- and oligonucleosomes in the cytoplasmic fraction of 125cell equivalents/well. |
| Specifications: | Assay time: 3 - 4 hours Negative control: Depending on cell culture conditions, each exponentially growing permanent cell culture contains a certain amount of dead cells (typically approximately 3 - 8%). In the immunoassay, these inherent dead cells in the untreated sample (without a cell-death-inducing agent) will cause a certain absorbance value (negative control). Positive control: A DNA-histone complex serves as a positive control. Sample material: Cytoplasmic fractions (lysates) of cell lines, cells ex vivo, cell culture supernatants, and serum or plasma |
| Specificity: | Anti-histone reacts with the histones H1, H2A, H2B, H3, and H4 of various species (e.g., human, mouse, rat, hamster, cow, opossum, Xenopus). Anti-DNA binds to single- and double-stranded DNA. Therefore, the ELISA allows the detection of mono- and oligonucleosomes from various species, and may be applied to measure apoptotic cell death in many different cell systems. |
| Unit Definition: | Unit Conversion: 1 mU = 1 x 10-3 OD (1 mU = 0.001 OD). |
| Symbol |    GHS07,GHS05,GHS07,GHS09 |
| Signal word | WarningDanger |
| Hazard statements | H317 - H319 - H412 - H317 - H318 - H411 |
| Precautionary statements | P261 - P273 - P280 - P261 - P273 - P280 - P333 + P313 - P337 + P313 - P362 + P364 - P305 + P351 + P338 + P310 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352200 |
| Components | Anti-histone-biotin antibody (clone H11-4); Anti-DNA-POD antibody (clone MCA-33); Positive Control; Incubation Buffer; Lysis Buffer; Substrate Buffer; ABTS Substrate Tablet; ABTS Stop Solution; Microplate (streptavidin-coated); Adhesive Cover Foils |