ApopTag® Peroxidase In Situ Apoptosis Detection Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Do not warm the stock reagents before dispensing them from containers. - Ethanol should not be substituted for n-butanol as it will destain nuclei.		- If staining of both nuclei and cytoplasm is apparent, the nonspecific component often can be mitigated by reducing the development time in substrate. - If a very high percentage of the nuclei in a tissue can be stained with TdT, but not without TdT, there appears to be staining of DNA only.

Upstream tips
- Do not warm the stock reagents before dispensing them from containers. - Ethanol should not be substituted for n-butanol as it will destain nuclei.
Downstream tips
- If staining of both nuclei and cytoplasm is apparent, the nonspecific component often can be mitigated by reducing the development time in substrate. - If a very high percentage of the nuclei in a tissue can be stained with TdT, but not without TdT, there appears to be staining of DNA only.

Publication protocol

The MetaMorph® microscopy automation and image analysis software was used for automatic nuclei counting for detection of apopototic cell bodies and cell proliferation (Universal Imaging Corporation, Downingtown, PA, USA). The ApopTag® peroxidase ISOL apoptosis detection kit (Millipore) was used on cultured cells or on paraffin sections from mouse tumor tissue. Apoptosis was also evaluated by counting the percentage of cells stained with DAPI that displayed chromatin condensation and nuclear fragmentation (apoptotic bodies). At least 500 cells were examined in each sample to quantify apoptosis. Parallel paraffin sections of mouse tumors were stained for Ki67 (Dako) to estimate cell proliferation.

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Manufacturer protocol

Download the product protocol from Millipore for ApopTag® Peroxidase In Situ Apoptosis Detection Kit below.

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