DNA Damage Assay Human Skin Fibroblast Cell (FSK)

DNA damage assay is a standard method for determining in-vivo/in-vitro genotoxicity by measuring the breaks in the DNA chain of animal and plant cells. Initial DNA damage leads to cell cycle arrest and, at the final stages, leads to induction of senescence or cell death (apoptosis, necrosis, autophagy, or mitotic catastrophe). Detection of DNA damage from mild to moderate to severe is challenging when studying genotoxicity in the pool of cells. It is favorable to use DNA damage assay kits available for prominent identification of the extent of damage in the analysis.

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Found 3 matching solutions for this experiment

CometAssay Electrophoresis System II

Bio-Techne

Upstream tips	Protocol tips	Downstream tips
Seed 2 × 10^4 cells/ml/gel	COMET assay included ketoprofen (KP, 10 μM) as positive control. Cells to be pretreated with KP at 37 °C in a CO2 incubator (100% relative humidity and dark conditions) for 30 minutes before the irradiation step.	The percentage of DNA damage of each sample could be calculated with the visual scoring of at least 100 DNA Comets using the following equation: [(Nclass 0 Comets × 0) + (Nclass 1 Comets × 1) + (Nclass 2 Comets × 2) + (Nclass 3 Comets × 3) + [(Nclass 4 Comets × 4) + (Nclass 5 Comets × 5) + (NClass 6 Comets × 6)]/6, where class 0 comets indicate comets with no DNA damage and class 6 comets indicate comets with maximum DNA damage.

Upstream tips
Seed 2 × 10^4 cells/ml/gel
Protocol tips
COMET assay included ketoprofen (KP, 10 μM) as positive control. Cells to be pretreated with KP at 37 °C in a CO2 incubator (100% relative humidity and dark conditions) for 30 minutes before the irradiation step.
Downstream tips
The percentage of DNA damage of each sample could be calculated with the visual scoring of at least 100 DNA Comets using the following equation: [(Nclass 0 Comets × 0) + (Nclass 1 Comets × 1) + (Nclass 2 Comets × 2) + (Nclass 3 Comets × 3) + [(Nclass 4 Comets × 4) + (Nclass 5 Comets × 5) + (NClass 6 Comets × 6)]/6, where class 0 comets indicate comets with no DNA damage and class 6 comets indicate comets with maximum DNA damage.

Manufacturer protocol Publication protocol 1 Relevant paper

H2A.X Phosphorylation Assay Kit (Flow Cytometry)

Sigma-Aldrich

Upstream tips	Protocol tips	Downstream tips
Seed 1×10^6 cells/ml on 145-mm dishes		Samples can be read on an LSR II flow cytometer and analyzed using Facsdiva software.

Upstream tips
Seed 1×10^6 cells/ml on 145-mm dishes
Downstream tips
Samples can be read on an LSR II flow cytometer and analyzed using Facsdiva software.

Publication protocol 1 Relevant paper

APO-BrdU™ TUNEL Assay Kit, with Alexa Fluor™ 488 Anti-BrdU

Thermo Fisher Scientific

Manufacturer protocol Publication protocol 1 Relevant paper

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