IN SITU CELL DEATH POD

General description

Kit for the detection and quantification of apoptotic cell death on a single-cell level by light microscopy in immunohistocytochemistry.

Application

The In Situ Cell Death Detection Kit, POD,  is a precise, fast, and simple nonradioactive technique to detect and quantify apoptotic cell death at the single-cell level in cells and tissues. Thus, the In Situ Cell Death Detection Kit can be used in many different assay systems.

• Examples are: Detection of individual apoptotic cells in frozen, paraffin-embedded and formalin-fixed tissue sections in basic research and routine pathology
• Determination of sensitivity of malignant cells to drug-induced apoptosis in cancer research and clinical oncology
• Typing of cells undergoing cell death in heterogeneous populations by double staining procedures

Features and Benefits

Sensitive: The maximum intensity of labeling (cell staining) of apoptotic cells is higher than the nick translation method.
Fast: The use of fluorescein-dUTP allows analysis of the samples directly after the TUNEL reaction, but before the addition of the secondary detection system.
Convenient: The direct labeling procedure using fluorescein-dUTP allows verification of the efficiency of the TUNEL reaction during the assay procedure.
Accurate: Identification of apoptosis at a molecular level (DNA-strand breaks) and identification of cells at the very early stages of apoptosis.
Flexible: No substrate included; provides the opportunity to select the staining procedure of choice.

Packaging

1 kit containing 3 components.

Principle

The In Situ Cell Death Detection Kit, POD is based on the detection of single- and double-stranded DNA breaks that occur at the early stages of apoptosis.
Apoptotic cells are fixed and permeabilized. Subsequently, the cells are incubated with the TUNEL reaction mixture that contains TdT and fluorescein-dUTP. During this incubation period, TdT catalyzes the addition of fluorescein-dUTP at free 3′-OH groups in single- and double-stranded DNA. After washing, the label incorporated at the damaged sites of the DNA is marked by an anti-fluorescein antibody conjugated with the reporter enzyme peroxidase. After washing to remove unbound enzyme conjugate, the POD retained in the immune complex is visualized by a substrate reaction.

Preparation Note

Storage conditions (working solution): TUNEL reaction mixture
The TUNEL reaction mixture should be prepared immediately before use and should not be stored. Keep TUNEL reaction mixture on ice until use.

Converter-peroxidase
Once thawed the Converter-peroxidase solution should be stored at 2 to 8 °C (maximum stability 6 months).
Note: Do not freeze.

Sample material: Cytospin and cell smear preparations, adherent cells grown on slides, and frozen and paraffin-embedded tissue sections.

Other Notes

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