MTX was from Sigma (St. Louis, MO). Human TNF-α was from BD Biosciences (Bedford, MA). KU-55933 and NU-7441 were from Tocris Biosciences. The following primary antibodies were used: mouse monoclonals: anti-actin (sc-8432; Santa Cruz Biotechnology), anti-P-ATM (NB110-55475; Novus), APC-anti-human CD3 (BD Biosciences), PE-anti-human CD14 (BD Biosciences); rabbit polyclonals: anti-JNK (sc-471; Santa Cruz Biotechnology), anti-p53 (NB200-171; Novus), anti-P-p65 (ab10684; Abcam), anti-p65 (ab7970; Abcam), anti-P-DNA-PKcs (ab18192; Abcam). The NF-κB-luciferase reporter (NF-κB-luc) plasmid with 5-κB elements was from Dean Ballard (Vanderbilt).
Anti p65 ab7970
Manufactured by Abcam
Anti-p65 (ab7970) is a primary antibody that detects p65, a subunit of the NF-κB transcription factor. This antibody can be used in various applications such as Western blotting, immunohistochemistry, and immunocytochemistry to study the expression and localization of p65.
Automatically generated - may contain errors
2 protocols using anti p65 ab7970
1
Investigating NF-κB Signaling Pathway
2
Chromatin Immunoprecipitation for NF-kB Binding
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RASFs were untreated or treated with LPS for 24 h before the cells were fixed with 1 % formaldehyde in medium for 10 minutes at room temperature. The sonication conditions were optimized to determine generation of DNA fragments between 300 and 600 bp in length. Briefly, nuclear lysates were sheared with five to eight pulses using an Ultrasonic Processor (80 W) with tubes immersed in iced water. Each pulse cycle comprised 5 s sonication followed by 25 s cooling down. Chromatin was immunoprecipitated with IgG (Sigma) and anti-p65 (ab7970, Abcam). The association of p50 and p65 was measured by RT-PCR on immunoprecipitated chromatin by use of the following primers: 5’-TATACTGGCTGTGACA-3’ (forward) and 5’-TGACAGATCCTGGCTT-3’ (reverse).
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