Hippocampal tissue was cleaved in a RIPA Lysis Buffer System that included protease inhibitors. After centrifugation (12,000 rpm, 20 min at 4 °C), the supernatant protein was measured using BCA protein assay reagent. Equal amounts of protein (30 μg) were separated with SDS–PAGE and transferred to PVDF membranes. The PVDF membranes were soaked in 5% skim milk for 1 h at room temperature and then incubated with primary anti-TLR4 antibody (1:1000 dilution), anti-p-p65 antibody (1:1000 dilution), anti-p65 antibody (1:1000 dilution), anti-p-IκBα antibody (1:1000 dilution), anti-IκBα antibody (1:1000 dilution), anti-Wnt3a antibody (1:1000 dilution) and anti-β-catenin antibody (1:1000 dilution) (Abcam, Cambridge, UK) at 4 °C overnight. After washing three times, the PVDF membranes were incubated with horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG secondary antibodies for 2 h at room temperature. Protein bands were detected using a chemiluminescence kit and visualized using Gel Imaging (Bio-Rad, Hercules, CA, USA).
Anti p iκbα antibody
Manufactured by Abcam
Sourced in United Kingdom
The Anti-p-IκBα antibody is a laboratory reagent used to detect the phosphorylated form of the IκBα protein. IκBα is a regulatory protein that inhibits the activity of the NF-κB transcription factor. Phosphorylation of IκBα leads to its degradation, allowing NF-κB to translocate to the nucleus and activate target genes. This antibody can be used to study the activation of the NF-κB signaling pathway.
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Lab products found in correlation
2 protocols using anti p iκbα antibody
1
Hippocampal TLR4 and Wnt3a Signaling
2
Protein Extraction and Immunoblot Analysis
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RIPA buffer (#P0013C, Beyotime, Shanghai, China) was used to isolate proteins. The lysates of cells or tissues were then centrifuged at 13,000 g for 15 mins to remove the debris. Next, the BCA assay (#P0012, Beyotime) was used to qualify the concentrations of proteins.
Primary antibodies including anti-USP8 antibody (#ab228572), anti-RIPK2 antibody (#ab8428), anti-p-IκBα antibody (#ab240059), anti-IκBα antibody (#ab133462), anti-iKKα antibody (#ab32041), anti-p-IKKα/β antibody (#ab194528), and anti-β-actin (#ab115777) were obtained from Abcam (Cambridge, MA). Secondary antibodies were purchased from Invitrogen. Immunoblot analysis was performed as previously described methods.
Primary antibodies including anti-USP8 antibody (#ab228572), anti-RIPK2 antibody (#ab8428), anti-p-IκBα antibody (#ab240059), anti-IκBα antibody (#ab133462), anti-iKKα antibody (#ab32041), anti-p-IKKα/β antibody (#ab194528), and anti-β-actin (#ab115777) were obtained from Abcam (Cambridge, MA). Secondary antibodies were purchased from Invitrogen. Immunoblot analysis was performed as previously described methods.
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