Uses
Samples prelabeled with BrdU are fixed with ethanol, then incubated with a monoclonal antibody to BrdU. The antibody to BrdU supplied with the kit contains an optimized mixture of nucleases. These nucleases generate single-stranded DNA fragments, which allow binding of the antibody to BrdU without destruction of the cellular morphology. A fluorescein-labeled antibody to mouse immunoglobulin is added, then bound to the anti-BrdU antibody. Subsequently, the sample is evaluated using an immunofluorescence microscope.
- Safe: No radioisotopes are used
- Easy to perform: Follows a standard immunofluorescence protocol
- Sensitive: Denaturation of DNA with nucleases allows for highly sensitive detection of BrdU
- Flexible: Allows double-labeling protocols
BrdU Labeling and Detection Kit has been used for the detection of 5-bromo-2′-deoxy-uridine (BrdU) incorporated into cellular DNA.
General Description
The kit is used for the detection of BrdU incorporated into cellular DNA using immunofluorescence microscopy. It is used for the detection of DNA synthesis by either in vitro labeling of cells or organ cultures, or by in vivo labeling, in which frozen or paraffin-embedded tissue sections must be prepared prior to fixation.
Normally, binding of the antibody is only achieved by denaturation of the DNA. This is usually obtained by exposing the cells to acid, base, or heat. These procedures result in destruction of cell integrity, including cell morphology and surface and cytoplasmatic markers.
The BrdU Labeling and Detection Kit I avoids these problems. The antibody preparation contains specific nucleases which allows access to BrdU after fixation in acidic ethanol. Therefore also simultaneous detection of other markers (double staining) is possible.
