Immunoblotting Analysis of Cell Signaling

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-p21^waf/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)].

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Conde M., Michen S., Wiedemuth R., Klink B., Schröck E., Schackert G, & Temme A. (2017). Chromosomal instability induced by increased BIRC5/Survivin levels affects tumorigenicity of glioma cells. BMC Cancer, 17, 889.

Publication 2017

AF886 AF1355 ab1431 monoclonal mouse anti-γH2AX

Actin Antibodies Cells Cyclin d1 Cyclin e Goat Membranes Mouse Rabbit Survivin

Corresponding Organization :

Other organizations : University Hospital Carl Gustav Carus, TU Dresden, Heidelberg University, German Cancer Research Center

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Variable analysis

independent variables

Treatments/Conditions (e.g., cell type, drug treatment)

dependent variables

Protein expression levels of Survivin, c-myc, p21^waf/cip, p53, p53 S15, Actin, ATM S1981, ATM, CHK2 T68, Cyclin D1, Cyclin E, γH2AX

control variables

Loading control (Actin)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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