DNase I recombinant Troubleshooting

Product No. 04536282001

Troubleshooting

Tissue sample thickness for efficient penetration of DNase I, grade I when using In Situ Cell Death Detection Kit

For the In Situ Cell Death Detection Kit (either AP or POD), the thickness of tissue sections should be up to 10 μm, or DNase may not be able to penetrate the slide efficiently. Test variations of the permeabilization step by empirically testing incubation times and incubation conditions.

Optimization the conditions of DNAse I, Grade I in TUNEL assays

For a TUNEL assay, dissolve DNase I in 50 mM Tris-HCl, pH 7.5, 1 mg/ml BSA. The final concentration will depend on the sample being processed. In general, start by using a low concentration (1 U/ml for 10 min at +15 to +25 °C). This procedure is described in the pack inserts of the In Situ Cell Death Detection Kits. Follow the steps described in the troubleshooting section of the pack inserts of the In Situ Cell Death Detection Kits when having problems with positive controls.

The pack inserts and the Apoptosis and Cell Proliferation Special Interest Site provide more information about the In Situ Cell Death Detection Kits. There are four types of Roche TUNEL Kits:

In Situ Cell Death Detection Kit, TMR red
In Situ Cell Death Detection Kit, Fluorescein
In Situ Cell Death Detection Kit, POD
In Situ Cell Death Detection Kit, AP

Materials

     
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