This product is intended as a positive control for use with all ApopTag In Situ Apoptosis Detection Kits. These reference slides are also useful as teaching aides, and for procedural troubleshooting. Each pack of ApopTag Control Slides contains 5 unstained positive control slides. Rat mammary glands were obtained at the fourth day after weaning and were fixed for 18 hours in 10% neutral buffered formalin. After embedding in paraffin, 5 mum thick sections were cut from the middle of the tissue and were mounted on silanized slides.
Application: Interpretation of Staining These positive control slides of female rat mammary tissue 4-days post weaning show only 1-2% of the cells being apoptotic. Microscopic inspection is usually required to confirm proper kit activity. Stain is generally localized to chromatin, and is intensely colored. The localization and sensitivity to be expected with the ApopTag Peroxidase (S7100, S7101) or ApopTag Fluorescein (S7110, S7111, S7160, S7165) Kits are similar. Positive objects vary in size, from intact nuclei to much smaller apoptotic bodies, and in shape, from round to irregular. Such objects may be either isolated, clustered, or engulfed in the cytoplasm of a phagocytic cell. Some unstained apoptotic nuclei or bodies may also be expected. Light cytoplasmic staining might co-localize with intense chromatin staining (this might be due to fragmented DNA leaching from the nucleus, or that in phagocytic organelles). If staining of non-apoptotic cell nuclei is observed with the ApopTag Peroxidase kits (S7100, S7101), the user can attempt to decrease the substrate development time or temperature, or to further dilute the anti-digoxigenin antibody reagent in 0.5% (w:v) BSA. Counterstaining is recommended (see The Complete ApopTag Manual). Limitations This control tissue has been selected for positive staining with all ApopTag Kits. Results with control tissue may vary from results with other tissues because of both sample preparation differences and intrinsic tissue differences.
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