anti egfr erb b1  (Bio-Techne corporation)

Journal: Molecular and Cellular Biology

Article Title: Phosphorylation of Chk1 by ATR Is Antagonized by a Chk1-Regulated Protein Phosphatase 2A Circuit

doi: 10.1128/mcb.00447-06

Figure Lengend Snippet: FIG. 3. Go¨6976 does not induce global phosphorylation of ATR targets or H2AX foci. HeLa cells were untreated (control) or were incubated with 20 mM HU (2 h), 20 M VP-16 (4 h), 1 M Go¨6976 (1 h), or 1 M UCN-01 (2 h). Cell lysates were resolved by SDS-PAGE and subjected to Western blotting with antibodies that recognize phosphorylated substrates of ATM/ATR (A) or phosphorylated Brca1 (B). -Catenin antibodies were used to verify equal loading. (C) HeLa cells were treated as in panels A and B, except that cells were incubated with Go¨6976 for 1 h, 2 h, 3 h, and 6 h before harvest. Samples were analyzed by Western blotting for pS1981-ATM, pS966-Smc1, pS345-Chk1, and Chk1. (D) HeLa cells were treated as in panel C and analyzed by indirect immunofluorescence for H2AX foci.

Article Snippet: Other primary antibodies used were anti-Cdc25A (Ab-3; Neomarkers), anti-phospho Smc1 S966 (Novus Biologicals), anti-phospho Brca1 S1423 (Chemicon), anti-PP1 (Upstate), anti-PP2A (Upstate), anti-PP4 (recognizes PP2A and PP4) (Abcam), anti-PP6 (Exalpha Biologicals), anti- -catenin (BD Transduction), anti- H2AX (Upstate), anti-ATR (Oncogene), and anti-p1981ATM (Rockland Biochemicals).

Techniques: Phospho-proteomics, Control, Incubation, SDS Page, Western Blot