H2A.X Phosphorylation Assay Kit (Flow Cytometry)

Protocol tips

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

In order to characterize the extent of DNA damage induced by chronic exposure to FA, FSK cells were seeded 1 × 10^6 on 145-mm dishes and treated for 24 h with 250 μM FA. Thereafter, cells were collected, washed in PBS and prepared for flow cytometry using the H2A.X Phosphorylation Assay Kit for flow cytometry (EMD Millipore) according to the manufacturer’s instructions. Briefly, cells were collected, stained and resuspended in 500 µl of PI in the dark for 30 min. Samples were read on an LSR II flow cytometer and analyzed using Facsdiva software. Each experiment was performed in duplicate, repeated at least three times using different cell batches.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for H2A.X Phosphorylation Assay Kit (Flow Cytometry) below.

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