In Situ Cell Death Detection Kit, Fluorescein

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Cells were treated with or without 5, 10 and 20 μM of HMJ-30 for 24 h		- Non specific labelling can be corrected by fixing cells by treating with 4% buffer PFA or Formalin. - When Nonspecific labelling is present, reduce concentration of TdT - When high background is present reduce concentration of labelling mix by 50%

Upstream tips
- Cells were treated with or without 5, 10 and 20 μM of HMJ-30 for 24 h
Downstream tips
- Non specific labelling can be corrected by fixing cells by treating with 4% buffer PFA or Formalin. - When Nonspecific labelling is present, reduce concentration of TdT - When high background is present reduce concentration of labelling mix by 50%

Publication protocol

Following treatment with 5, 10 and 20 μM of HMJ-30 for 24 h or without treatment, apoptosis in the HUVECs was determined by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, using an In Situ Cell Death Detection kit, fluorescein (Roche Diagnostics GmbH, Mannheim, Germany) and flow cytometric analysis. Following HMJ-30 exposure, cells were prepared for detecttion of DNA fragmentation following a previously reported method (32,33).

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Manufacturer protocol

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