p-γH2AX immunohistochemistry assay

LH Lidia F. Hernandez
AD Angie B. Dull
SK Soumya Korrapati
CA Christina M. Annunziata
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Ovcar8 cells expressing WT or low Caspase 8 levels were subjected to treatment with carboplatin, birinapant, or combined simultaneous treatment with carboplatin and antagonistic dose of birinapant for 18 h. Cells were fixed with 4% paraformaldehyde and standard immunohistochemistry was performed to assess the levels of phosphorylated γH2AX as a measure of carboplatin-induced DNA damage. Subcutaneous xenograft tumors from mice treated with single, simultaneous, and sequential drug combinations were freshly fixed after a week of treatment and subjected to phosphorylated γH2AX level assessment using the same antibody (p-γH2AX antibody Millipore # 05-636-AF488). Experiment included ten mice per group.

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