MERCK/Millipore
Anti-phospho-Histone H2A.X (Ser139) Antibody
05-636-I
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In response to strand breaks in the DNA, ATM phosphorylates H2AX on serine 139, which is required for DNA damage signaling and repair. Our aim is to examine the levels of phospho-Histone H2A.X (Ser139) in HeLa cells treated with bleomycin (5 µM) for 2 hours. Bleomycin induces double-strand breaks in the DNA. Histones were prepared from HeLa cells by acid precipitation method, and we analyzed the levels H2A.X (Ser139) by western blot.
Western Blot
HeLa histones
1:2000, Overnight at 4°C
5% skimmed milk, 1 hr at room temperature
1:10000, anti-rabbit HRP conjugated antibody, 1 hr at room temperature
N/A
ECL reagent, Chemi doc
We observed a single specific band at 17 kDa corresponding to H2AX size in bleomycin-treated cells suggesting the induction of H2AX-ser139 in bleomycin-treated cells compared with untreated cells.
Excellent antibody for performing western blots
Good antibody for western blots and easy to optimise in mammalian cells.