Detection of Human Phospho RelA (S529) by Western Blot.  Samples: Whole cell lysate (50 ug for WB; 1 mg for IP, 20% of IP loaded) from Jurkat cells that were treated with TNF alpha and Calyculin A (+) or mock treated (-).   Antibodies: Affinity purified rabbit anti-Phospho RelA (S529) antibody A302-063A used for WB at 0.1 ug/ml (A) and 1 ug/ml (B).  In B, RelA was immunoprecipitated with affinity purified rabbit anti-RelA antibody A301-824A.  Immunoprecipitated RelA was either treated with phosphatase or mock treated.  Detection: Chemiluminescence with exposure times of 30 seconds (A and B).