Total protein was extracted using radioimmunoprecipitation assay buffer comprising protease and phosphatase inhibitors (Beyotime Biotechnology, Shanghai, China). Nuclear and cytoplasmic proteins were extracted with Nuclear and Cytoplasmic Protein Extraction Kit (KeyGen Biotech, Nanjing, China). Proteins were quantified using the bicinchoninic acid method based on the manufacturer’s agreement (Beyotime Biotechnology). The following primary antibodies were used in this study: anti-CDX2 (1:1,000; Cell Signaling Technology, #12306), anti-MUC2 (1:1,000; Abcam, ab134119), anti-KLF4 (1:1,000; Abcam, ab215036), anti-GATA4 (1:100; Santa Cruz Biotechnology, sc-25310), anti-NF-κB p105/p50 (1:1,000; Abcam, ab32360), anti-NF-κB p65 (1:1,000; Cell Signaling Technology, #4764), anti-phospho-NF-κB p50 (1:1,000; Abmart, PA1798), anti-phospho-NF-κB p65 (1:1,000; Cell Signaling Technology, #3033) and anti-Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:1,000; ZSGB-BIO, TA08). Goat anti-rabbit IgG and anti-mouse IgG (Biosharp) was used as secondary antibodies. Protein bands were visualized using chemifluorescence.
Anti nf κb p105 p50
Manufactured by Abcam
Anti-NF-κB p105/p50 is a primary antibody that recognizes the p105/p50 subunit of the NF-κB transcription factor. NF-κB is a key regulator of immune and inflammatory responses.
Automatically generated - may contain errors
Lab products found in correlation
Anti nf κb p65, by Cell Signaling Technology (2 mentions)
Anti klf4, by Abcam (1 mentions)
Anti muc2, by Abcam (1 mentions)
Anti cdx2, by Cell Signaling Technology (1 mentions)
Anti phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Anti gata4, by Santa Cruz Biotechnology (1 mentions)
Nuclear and cytoplasmic protein extraction kit, by Keygen Biotech (1 mentions)
Radioimmunoprecipitation assay buffer, by Beyotime (1 mentions)
Protease and phosphatase inhibitor, by Beyotime (1 mentions)
Anti lamin a c, by Santa Cruz Biotechnology (1 mentions)
Anti p ikkα β, by Cell Signaling Technology (1 mentions)
Anti bcl 3, by Santa Cruz Biotechnology (1 mentions)
Anti p p38, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Santa Cruz Biotechnology (1 mentions)
Anti p akt, by Cell Signaling Technology (1 mentions)
Anti p iκbα, by Cell Signaling Technology (1 mentions)
Anti tak1, by Cell Signaling Technology (1 mentions)
Anti p85, by Merck Group (1 mentions)
Anti tubulin, by Merck Group (1 mentions)
Anti p38, by Santa Cruz Biotechnology (1 mentions)
2 protocols using anti nf κb p105 p50
1
Protein Extraction and Western Blot Analysis
2
Western Blot Analysis of Cell Signaling
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Cells were pelleted and solubilized with 0.1% Na-deoxycholate and 0.5% NP-40 in phosphorylation solubilisation buffer (PSB) at 4 °C51 (link). Whole-cell lysates were subjected to SDS-PAGE and Western blot analysis. The Western blot antibodies anti-BCL3 (rabbit polyclonal, clone C-14, sc-185), anti-GAPDH (mouse monoclonal, clone 6C5, sc-32233), anti-Lamin A/C (goat polyclonal, clone N-18, sc-6215), and anti-p38 (rabbit polyclonal, clone C-20, sc-535) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The antibodies anti-P-p38 (anti-phosphoThr180/phosphoTyr182, mouse monoclonal, clone 28B10, #9216), anti-P-AKT (anti-phosphoSer473, mouse monoclonal, clone 587F11, #4051), anti-TAK1 (rabbit monoclonal, clone D94D7, #5206), anti-P-IκBα (anti-phosphoSer32, mouse monoclonal, clone 14D4, #2859), anti-NFκB p65 (rabbit monoclonal, clone C22B4, #4764), anti-P-IKK-α/-β (anti-phosphoSer176/180, rabbit monoclonal, clone 16A6, #2697), and anti-IκBα (rabbit polyclonal, #9242) were obtained from Cell Signaling Technologies (Danvers, MA, USA) and the antibody anti-p85 (rabbit polyclonal, #06-195) was acquired from Millipore (Billerica, MA, USA). The antibody anti-Tubulin (mouse monoclonal, clone B512, #T5168) was purchased from Sigma-Aldrich (St. Louis, MO, USA) and the antibody anti-NFκB p105/p50 (rabbit polyclonal, ab7971, #GR40002-2) from Abcam (Cambridge, UK).
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