Mouse monoclonal anti histone h3

We performed western blot analysis of FLAG expression and NF-κB signaling in cell pellets harvested from ascites from control and IKFM ID8-Luc-injected C57BL/6 mice. Subcellular fractionation, western blotting, and signal detection were performed as previously described [34 (link)]. Primary antibodies used were rabbit polyclonal anti-FLAG (Abcam; 1:100) and rabbit polyclonal anti-p65 (Cell Signaling Technology; 1:1000). We confirmed the efficiency of fractionation using mouse monoclonal anti-histone H3 (Cell Signaling Technology; nuclear specific marker; 1:1000 dilution) and mouse monoclonal anti-α-tubulin (Sigma Chemical Co.; cytoplasmic specific marker; 1:1000 dilution). All original, uncropped gels are available in Additional file 3.

Protein isolated from ascites cells in the TBR5 experimental model was used for western blot analysis of IκBα expression. Whole protein isolation, western blotting, and signal detection were performed as previously described [57 (link)]. Primary antibodies used were rabbit polyclonal anti-IκBα (1:100 dilution, Cell Signaling Technology; 9242). Equal loading was confirmed using mouse monoclonal anti-histone H3 (1:1000 dilution, Cell Signaling Technology; 14269S) as a loading control. To image loading control and experimental antibody on the same blot, the gel was cut prior to hybridization. The pieces of the uncropped blot are shown in Supplemental Fig. S12.