primary antibodies against nf κb p65  (Cell Signaling Technology Inc)

Journal: Scientific Reports

Article Title: 3-carbamoyl proxyl nitroxide attenuates CCl 4 -induced liver fibrosis in mice through antioxidant-inflammatory regulation of TLR4/NF-κB signaling pathway

doi: 10.1038/s41598-026-46137-1

Figure Lengend Snippet: Molecular docking analysis of 3-CP with TLR4 and p65. ( A - B ) Predicted 3D binding model of 3-CP and TLR4/p65. 3-CP is colored purple. The protein surfaces of TLR4/p65 are colored in cyan. Potential interactions are indicated by dashed lines. ( C ) Predicted protein-ligand binding affinities (kcal/mol).

Article Snippet: Primary antibodies against NF-κB p65 (Cat #8242S), α-SMA (Cat# 19245 S), collagen I (Cat# 72026 S) and secondary antibody anti-rabbit IgG (CST#7074S) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Binding Assay, Ligand Binding Assay

Journal: Scientific Reports

Article Title: 3-carbamoyl proxyl nitroxide attenuates CCl 4 -induced liver fibrosis in mice through antioxidant-inflammatory regulation of TLR4/NF-κB signaling pathway

doi: 10.1038/s41598-026-46137-1

Figure Lengend Snippet: 3-CP is associated with inhibition of the TLR4/NF-κB pathway in the CCl 4 -induced liver fibrosis mice model. ( A - B ) Protein expression levels of TLR4/NF-κB signaling-associated proteins (TLR4, MyD88, IKKβ, p65/p-p65 and IκBα/p-IκBα). ( B - C ) Representative results of immunofluorescence staining for p-p65 and p-IκBα (scale bar: 50 μm). ( D ) Quantification of TLR4, MyD88, IKKβ, p65/p-p65 and IκBα/p-IκBα protein expression. Data are presented as mean ± SD, n = 3. * p < 0.05, vs. control group; # p < 0.05, vs. model group.

Article Snippet: Primary antibodies against NF-κB p65 (Cat #8242S), α-SMA (Cat# 19245 S), collagen I (Cat# 72026 S) and secondary antibody anti-rabbit IgG (CST#7074S) were purchased from Cell Signaling Technology (Danvers, MA, United States).

Techniques: Inhibition, Expressing, Immunofluorescence, Staining, Control

Journal: Redox Biology

Article Title: Ultrasmall Cu 2−x Se nanoparticles alleviate vascular calcification through inhibiting oxidative stress and NF-κB/NLRP3-mediated inflammation

doi: 10.1016/j.redox.2025.103961

Figure Lengend Snippet: CSP NPs inhibit NF-κB/NLRP3 signaling pathway during vascular calcification. Rat vascular smooth muscle cells (VSMCs) were treated with growth medium (GM), calcifying medium (CM), or CM with CSP NPs (2, 5, and 10 μg/mL) for 7 days (n = 4). (A) Representative western blots of p-p65 and p65. (B) Quantification of p-p65 protein expression by densitometry. Rat VSMCs were treated with GM, CM, or CM with CSP NPs (10 μg/mL) for 7 days (n = 4). (C) Representative immunostaining images of p65. Nuclear localization of p65 was determined by confocal microscopy. Scale bar = 25 μm. Rat VSMCs were treated with GM, CM, or CM with CSP NPs (2, 5, and 10 μg/mL) for 7 days (n = 4). (D) Representative western blots of NLRP3, Cleaved-Caspase1, IL-1β, and IL-6. (E) Quantification of NLRP3, Cleaved-Caspase1, IL-1β, and IL-6 protein expression by densitometry. VitD 3 -overloaded mice were treated with CSP NPs (1 mg/kg) for 8 days (n = 6). (F) Representative western blots of p-p65, p65, and NLRP3. (G) Quantification of p-p65 and NLRP3 protein expression by densitometry. Rats subjected to 5/6 nephrectomy were treated with CSP NPs (0.7 mg/kg) for 4 weeks (n = 6). (H) Representative western blots of p-p65, p65, and NLRP3. (I) Quantification of p-p65 and NLRP3 protein expression by densitometry. ∗ P< 0.05, ∗∗ P< 0.01.

Article Snippet: Immunostaining of cells was performed by overnight incubation at 4 °C with NF-κB p65 primary antibody (8242S, Cell Signaling Technology; 1:400 dilution in BSA/PBS), followed by 60-min incubation with Alexa Fluor 488-conjugated secondary antibodies (1:200), and nuclear counterstaining with Hoechst 33342.

Techniques: Western Blot, Expressing, Immunostaining, Confocal Microscopy

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: A novel peptide-compound conjugate alleviates endotoxin-induced inflammation via NF-κB/MAPK modulation

doi: 10.1007/s00109-025-02608-y

Figure Lengend Snippet: a Effects of 2IP5MP-CW on the expression of phosphorylated IKKβ, IkBα, and NF-kB protein in LPS-induced RAW 264.7 cells. ### P < 0.001 compared with the control group. *** P < 0.001 compared with LPS group. All experiments were performed in triplicate. b The 2IP5MP-CW blocks LPS-induced NF-κB nuclear translocation, as demonstrated by the decreased green fluorescence of phosphorylated NF-κB p65, with the nuclei counterstained in blue by DAPI. The graph shows a quantitative analysis of p-NF-κB p65 levels, indicating a significant reduction in NF-κB activation upon treatment with 2IP5MP-CW. ### P < 0.001 compared with the control group. *** P < 0.001 compared with LPS group

Article Snippet: Non-specific binding was blocked with 5% bovine serum albumin for 1 h. Cells were incubated overnight at 4 °C with rabbit anti-phospho-NF-κB p65 primary antibody (Cell Signaling Technology, Danvers, MA, USA) and subsequently with goat anti-rabbit IgG Texas Red conjugate (1:500, Santa Cruz Biotechnology, Dallas, TX, USA) for 3 h at room temperature.

Techniques: Expressing, Control, Translocation Assay, Fluorescence, Activation Assay