abs raised against p-nf-κb p65 antibody  (Cell Signaling Technology Inc)

Journal: Science Advances

Article Title: PmiR senses 2-methylisocitrate levels to regulate bacterial virulence in Pseudomonas aeruginosa

doi: 10.1126/sciadv.add4220

Figure Lengend Snippet: ( A to C ) Inflammatory cytokines in BALF were assessed by enzyme-linked immunosorbent assay. ( D ) Western blot detected the expression of inflammatory cytokines (TNF-α, IL-1β, and IL-6), pyroptosis markers (caspase-1 and GSDMD), STAT3, p-STAT3, NF-кB/p65, and p-NF-кB/p65. ( E ) The densitometry of the data shown in (D). Error bars indicate the SD from three independent replicates. Groups were compared with each other in statistics. * P < 0.05, *** P < 0.001, **** P < 0.0001 based on one-way ANOVA Dunnett’s multiple comparison test.

Article Snippet: The Abs were used from different vendors as stated below: mouse Abs against p-NF-κB/p65 (sc-136548), caspase-1 (sc-514), TNF-α (sc52746), IL-6 (28343), and β-actin (sc-47778) were obtained from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Comparison

Journal: Frontiers in immunology

Article Title: oprC Impairs Host Defense by Increasing the Quorum-Sensing-Mediated Virulence of Pseudomonas aeruginosa .

doi: 10.3389/fimmu.2020.01696

Figure Lengend Snippet: FIGURE 5 | oprC deficiency decreased pyroptosis and STAT3/ NF-κB phosphorylation. (A) MH-S cells challenged with bacteria at a multiplicity of infection (MOI) of 10 for 2 h. Immunoblotting analysis of GSDMD, NF-κB/p65, p-NF-κB/p65, STAT3, and p-STAT3. (B,C) Secreted IL-1β and LDH from the supernatant was assessed by ELISA and LDH assay kit after MH-S cells were infected at an MOI of 10 for 2 h (n = 3). (D) Representative images of immunofluorescence staining of the lungs infected with bacteria for p-STAT3 co-stained with p-NF-κB/p65 and DAPI (n = 3). Arrows indicate the colocalizations between p-STAT3 and p-NF-κB/p65. Scale bars, 10 µm. Error bars represent the mean ± s.d. One-way ANOVA with a post-hoc Tukey test was performed for comparison of means of groups.

Article Snippet: Mouse Abs against p-p65 (p-NFκB p65 Antibody [Ser 536]: sc-136548), ASC (ASC Antibody [B-3]: sc-514414), caspase1(caspase-1 p10 Antibody [M-20]: sc-514), and β-Actin (β-Actin Antibody [C4]: sc-47778) were obtained from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Phospho-proteomics, Bacteria, Infection, Western Blot, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay, Staining, Comparison

Journal: Frontiers in immunology

Article Title: oprC Impairs Host Defense by Increasing the Quorum-Sensing-Mediated Virulence of Pseudomonas aeruginosa .

doi: 10.3389/fimmu.2020.01696

Figure Lengend Snippet: FIGURE 6 | STAT3/NF-κB inhibitors decreased pyroptosis following P. aeruginosa infection. MH-S cells pretreated with 10 µm stattic (the inhibitor of Stat3 activation) for 30 min, or BAY (the inhibitor of NF kappa B activation) for 1 h, were challenged with bacteria at a multiplicity of infection (MOI) of 10 for 2 h. Cell lysis and supernatants were used for western blot and ELISA analysis, respectively. (A,B) Western blot analysis of STAT3, p-STAT3, NF-κB/p65, p-NF-κB/p65, and GSDMD. (C–H) TNF-α, IL-1β, and IL-6 were assessed by ELISA (n = 3). Error bars represent the mean ± s.d. One-way ANOVA with a post-hoc Tukey test was performed for comparison of means of groups.

Article Snippet: Mouse Abs against p-p65 (p-NFκB p65 Antibody [Ser 536]: sc-136548), ASC (ASC Antibody [B-3]: sc-514414), caspase1(caspase-1 p10 Antibody [M-20]: sc-514), and β-Actin (β-Actin Antibody [C4]: sc-47778) were obtained from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Infection, Activation Assay, Bacteria, Lysis, Western Blot, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Effective Pro-Inflammatory Induced Activity of GALT, a Conserved Antigen in A. Pleuropneumoniae , Improves the Cytokines Secretion of Macrophage via p38, ERK1/2 and JNK MAPKs Signal Pathway

doi: 10.3389/fcimb.2018.00337

Figure Lengend Snippet: Analysis of the recognition receptor of GALT. Anti-TLR2 and anti-TLR4 antibodies were used to block the corresponding Toll-like receptor. Secretion of pro-inflammatory cytokines were detected to analyze the function of TLR2 and TLR4.

Article Snippet: Subsequently, these proteins were probed with specific Abs against P38, P-P38, Erk l/2, p-Erk l/2, JNK, P-JNK, P65, P-P65, TLR2, and TLR4 (Cell Signaling Technology).

Techniques: Blocking Assay

Journal: Innate immunity

Article Title: A novel nucleotide oligomerisation domain 2 mutation in a family with Blau syndrome: Phenotype and function.

doi: 10.1177/1753425917727063

Figure Lengend Snippet: Figure 2. Patients with NOD2 E383D have decreased NF-kB activation and increased levels of P-ERK. Equal numbers of freshly isolated PBMCs from healthy donors (C1–C3), patients with BS (P1–P3) and treatment controls (T1 and T2) were lysed and loaded onto SDS-PAGE. This was followed by Western blot for activation markers of NF-kB and MAP kinase. Levels of P-p65 and P-IkBa are decreased in patient samples and treatment controls vs. healthy controls. Levels of P-p38 were similar between patients and controls. Total GAPDH levels act as a loading control. The figure represents a single Western blot experiment.

Article Snippet: Abs against P-p65 (#3033), IkBa (#9242), P-IkBa (#9246), P-ERK (#9101) and P-p38 (#4511) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Activation Assay, Isolation, SDS Page, Western Blot, Control