Anti NFkB P52  

EMSA results using Anti-NFKB p52. Comparison of nuclear factor (NF)-κB binding activity among CD30-positive lymphomas with or without stimulation of CD30. (a) Whole-cell lysates of CD30-positive lymphoma cell lines with or without stimulation of CD30 were incubated with the？32P-labeled HIV κB probe, and κB binding was analyzed by electrophoretic mobility shift assay (EMSA). (b) Supershift assay of CD30-stimulated Karpas 299 and SR-786 using the antibodies indicated. Addition of anti-Bcl-3 antibodies produced no apparent supershifted bands (data not shown). The specificity of κB binding was determined by competition with an excess amount of unlabeled HIV κB probe or control unlabeled AP1 probe (5′-CGCTTGATGAGTCAGCCGGAA-3′). (c) NF-κB DNA-binding activities of p52-containing components were compared among CD30-positive lymphoma cell lines upon stimulation of CD30. The whole-cell lysates were incubated with the？32P-labeled H2 κB probe, and the binding was analyzed by EMSA. (d) Supershift assay of CD30-stimulated Karpas 299, KM-H2 and KIS-1. The cell lysates were first incubated with antibodies against p50, p52, p65, RelB and c-Rel, and the components of the NF-κB-binding complexes in each cell line were determined. The specificity of κB binding was determined by competition with an excess amount of unlabeled H2 κB probe or control unlabeled AP1 probe. Fig 5. PMID: 16108830.