antinfκb p65 antibody  (MyBiosource Biotechnology)

Journal: SLAS discovery : advancing life sciences R & D

Article Title: A Screening Approach for Identifying Gliadin Neutralizing Antibodies on Epithelial Intestinal Caco-2 Cells.

doi: 10.1177/2472555217697435

Figure Lengend Snippet: Figure 4. NF-κB activation by PT-gliadin. Caco-2 monolayers were challenged with PT-gliadin (1 mg/mL) or with preincubated solutions of PT-gliadin and anti-PT-gliadin IgY (3.5 mg/mL), or were left untreated for 45 min. (A) Whole cell lysates were subjected to Western blot analysis for phosphorylated NF- κB p65, total NF-κB p65, and GAPDH (as a loading control). Changes of (B) phosphorylated NF-κB p65 and total NF-κB p65 levels upon addition of anti-PT-gliadin IgY (specific IgY) were calculated by normalizing data to the PT-gliadin-treated sample (= 1). Data are depicted as means ± SEM; *indicates a significant difference with p < 0.05.

Article Snippet: The gels were blotted to nitrocellulose membranes (TransBlot Turbo Mini Nitrocellulose Transfer Packs, Bio-Rad), and the membranes blocked afterward with 5% BSA in TBS-T (50 mM Tris [pH 7.5], 150 mM NaCl, and 0.1% Tween-20) and probed with the primary antibodies antiNFκB p65 (sc-8008, Santa Cruz), anti-phospho-NFκB p65 (Ser536; 3033S, CST), and anti-GAPDH (2118S, CST) as control.

Techniques: Activation Assay, Western Blot, Control