CELL DEATH DETECTION ELISA

General description

The Cell Death Detection ELISA serves as photometric enzyme immunoassay for the qualitative and quantitative in vitro determination of cytoplasmic histone-associated DNA fragments (mono- and oligonucleosomes) after induced cell death. This assay is based on the quantitative sandwich-enzyme immunoassay-principle using mouse monoclonal antibodies directed against DNA and histones, respectively. The antibodies used are not species specific.

Specificity

Anti-histone antibody reacts with the histones H1, H2A, H2B, H3, and H4 of various species (e.g., human, mouse, rat, hamster, cow, opossum, and Xenopus). Anti-DNA-POD antibody binds to ss- and dsDNA. 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.

Application

Cell Death Detection ELISA has been used to measure Caspase-3 activity, intracellular reactive oxygen species (ROS) and cell apoptosis.

For research use only. Not for use in diagnostic procedures.
Specific determination of mono- and oligonucleosomes in the cytoplasmic fraction of cell lysates.

Packaging

1 kit containing 9 components.

Preparation Note

Working solution: Coating solution
Predilute 1 ml Coating buffer conc. with 9 ml double dist. water. Shortly before use, dilute 1 ml anti-histone antibody (reconstituted) with 9 ml Coating buffer.

Washing solution
Warm the Washing buffer concentrate to 15 to 25 °C and dilute 40 ml in 360 ml double distilled water. Mix thoroughly.

Sample solution
Preparation of the sample solution depends on the cell system used and the extent of cell death.
Example: Dilute 25 μl of sample in 225 μl Incubation buffer.

Conjugate solution
Dilute 1 ml Anti-DNA-POD (reconstituted) with 9 ml Incubation buffer.

Substrate solution
Depending on the number of samples tested, dissolve 1, 2, or 3 tablets in 5, 10, or 15 ml Substrate buffer.
Allow to come to 15 to 25 °C before use.
Note: The ABTS solution is light sensitive over extended periods of time.
Storage conditions (working solution): Anti-histone
2 to 8 °C for 2 months
Anti-DNA-POD
2 to 8 °C for 2 months
Coating solution
Prepare immediately before use!
Washing solution
2 to 8 °C for 2 months
Sample solution
2 to 8 °C for 2 months
Conjugate solution
Prepare immediately before use!
Substrate solution
2 to 8 °C for 1 month, protected from light.

Reconstitution

Anti-histone
Reconstitute the lyophilizate in 1 ml double-distilled water for 10 minutes.Mix thoroughly.
Anti-DNA-POD
Reconstitute the lyophilizate in 1 ml double-distilled water for 10 minutes. Mix thoroughly.

Other Notes

For life science research only. Not for use in diagnostic procedures.