Total cells lysates were prepared and analyzed by immunoblotting as described previously [40 (link)]. Membranes were incubated with primary antibodies including polyclonal anti-Survivin (AF886, R&D Systems), monoclonal anti c-myc (R950–25, Invitrogen), monoclonal rabbit anti-p21waf/cip (#2947, Cell Signaling), polyclonal goat anti-p53 (AF1355, R&D Systems), monoclonal rabbit anti-p53 S15 (ab1431, Abcam), monoclonal mouse anti-actin (A2228, Sigma), monoclonal rabbit anti-ATM S1981 (#2152–1, Epitomics), polyclonal rabbit anti-ATM (PC116, Merck, Darmstadt, Germany), monoclonal rabbit anti-CHK2 T68 (#2661, Cell Signaling), polyclonal rabbit anti-Cyclin D1 (sc-753, Santa Cruz), polyclonal rabbit anti-Cyclin E (sc-247, Santa Cruz), and monoclonal mouse anti-γH2AX (05–636, Millipore). Bound antibodies were detected using appropriate secondary antibodies conjugated with HRP (Dako, Hamburg, Germany) as described previously [40 (link)].
Monoclonal mouse anti γh2ax
Manufactured by Merck Group
Sourced in Germany, United States
Monoclonal mouse anti-γH2AX is a laboratory reagent used for the detection of phosphorylated histone H2AX (γH2AX), a marker of DNA double-strand breaks. It is an antibody that specifically binds to the phosphorylated form of the H2AX histone protein.
Automatically generated - may contain errors
2 protocols using monoclonal mouse anti γh2ax
1
Immunoblotting Analysis of Cell Signaling
2
Immunocytochemistry of γH2AX in BTC Cells
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Immunocytochemistry studies of γH2AX were performed using BTC cells. As a positive control, we also assessed BTC cells 4 h after 6 Gray irradiation using a Gamma Cell 40 Exactor (Nordion International, Ottawa, ON, Canada). Briefly, cells were cultured on six-well chamber slides, fixed with 4% paraformaldehyde, and permeabilized. The cells were then incubated with monoclonal mouse anti-γH2AX (diluted 1:500 [Millipore, Billerica, MA, USA]), followed by Alexa Fluor anti-mouse IgG conjugated to Alexa Fluor 488 (diluted 1:500 [Cell Signaling Technology, Danvers, MA, USA]). The slides were viewed by fluorescence microscopy (BZ-8000 [Keyence, Osaka, Japan]).
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