mouse atm antibody  (Novus Biologicals)

Journal: Oncotarget

Article Title: Inhibition of RAC1 GTPase sensitizes pancreatic cancer cells to γ-irradiation.

doi: 10.18632/oncotarget.2500

Figure Lengend Snippet: Figure 5: Rac1 inhibition abolishes IR-induced activation of both ATM and ATR signaling pathways. CD18/HPAF cells were treated with/without 10 Gy IR in the presence of NSC23766 at the indicated doses and incubated for 1 h at 37oC. (A) To assess ATR and ATM kinase activities, ATR and ATM were immunoprecipitated from the cell lysates using anti-ATR (N-19) and anti-ATM (2C1) antibodies respectively and assayed for relative kinase activity using recombinant p53 protein as substrate. (B) To measure Chk1 and Chk2 activity, Chk1 and Chk2 were immunoprecipitated from the cell lysates using anti-Chk1 (G-4) and anti-Chk2 (B-4) antibodies respectively and assayed for relative kinase activity using recombinant Cdc25C protein as substrate. As controls, protein levels of ATR, ATM, Chk1 and Chk2 in the immunoprecipitates (IP-WB) as well as in the cell lysates (WB) were assessed by immunoblotting. (C) Cdc2 was immunoprecipiated from the cell lysates using anti-Cdc2 (17) antibody and analyzed for Cdc2-Y15 phosphorylation and Cdc2 protein by immunoblotting.

Article Snippet: These included mouse IgG for ATM (2C1) (Novus Biologicals, Littleton, CO), Cdc2 (17), Chk1 (G-4), Chk2 (B-4) and poly(ADP-ribose) (3H2844); rabbit IgG for ATM (Ab-3) (EMD Biosciences, San Jose, CA), Caspase 3 (Cell Signaling, Danvers, MA), Cdc2 (C-19), Chk1 (FL-476), Chk2 (Cell Signaling, Danvers, MA), GAPDH (FL-335), Rac1 (C-14); and goat IgG for Actin (I-19), ATR (N-19) and phospho-Cdc2 (Tyr15).

Techniques: Inhibition, Activation Assay, Protein-Protein interactions, Incubation, Immunoprecipitation, Activity Assay, Recombinant, Western Blot, Phospho-proteomics