pstat3-pe antibody ebioscience #501122408  (Thermo Fisher)

Journal: Scientific Reports

Article Title: Unveiling novel insights into human IL-6 − IL-6R interaction sites through 3D computer-guided docking and systematic site mutagenesis

doi: 10.1038/s41598-024-69429-w

Figure Lengend Snippet: Activation of phosphorylation of STAT3 in HepG2 cells by hIL-6 or its mutants. HepG2 cells were treated with 50 ng/mL hIL-6 or its mutants for 15 min, and cell lysates were analyzed to detect phosphorylation of STAT3. Untreated cells were used as the negative control (NC). ( A ) Representative, digital western blot images of expression of P-STAT3, STAT3 and β-Actin upon stimulation with hIL-6 WT or R167A, E171A and R178E mutants, respectively; ( B ) statistical analysis of hIL-6 mutants compared with hIL-6 WT in term of STAT3 phosphorylation levels in HepG2 cells, based on three independent experiments. Data is presented as mean ± SD, n = 3. **p < 0.01; and ***p < 0.001.

Article Snippet: The antibody, PE labeled anti-pSTAT3 (pY705) monoclonal antibody was purchased from BD (Cat. 612569), and horseradish peroxidase (HRP)-conjugated goat anti-human IgG antibody was obtained from Southern Biotech (Cat. 2049–5).

Techniques: Activation Assay, Negative Control, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Unveiling novel insights into human IL-6 − IL-6R interaction sites through 3D computer-guided docking and systematic site mutagenesis

doi: 10.1038/s41598-024-69429-w

Figure Lengend Snippet: Activate of phosphorylation of STAT3 in Leukocytes by IL-6 mutants. Human leukocytes were preincubated with 50 ng/mL IL-6 mutants for 15 min, followed by permeabilized and stained. Untreated cells were used as the negative control (NC). Phosphorylation of STAT3 was assessed using flow cytometry with PE labeled anti-pSTAT3 (pY705) monoclonal antibody. ( A – C ) Gating strategy and representative histogram graph of P-STAT3 + cells.; ( D ) statistical analysis of percentage of P-STAT3 + cells upon stimulation with hIL-6 WT or R167A, E171A and R178E mutants, based on three independent experiments. The data is presented as mean ± SD, n = 3. *p < 0.05; **p < 0.01.

Article Snippet: The antibody, PE labeled anti-pSTAT3 (pY705) monoclonal antibody was purchased from BD (Cat. 612569), and horseradish peroxidase (HRP)-conjugated goat anti-human IgG antibody was obtained from Southern Biotech (Cat. 2049–5).

Techniques: Staining, Negative Control, Flow Cytometry, Labeling

Journal: Scientific Reports

Article Title: Unveiling novel insights into human IL-6 − IL-6R interaction sites through 3D computer-guided docking and systematic site mutagenesis

doi: 10.1038/s41598-024-69429-w

Figure Lengend Snippet: Activation of phosphorylation of STAT3 in HepG2 cells by hIL-6 or its mutants. HepG2 cells were treated with 50 ng/mL hIL-6 or its mutants for 15 min, and cell lysates were analyzed to detect phosphorylation of STAT3. Untreated cells were used as the negative control (NC). ( A ) Representative, digital western blot images of expression of P-STAT3, STAT3 and β-Actin upon stimulation with hIL-6 WT or R167A, E171A and R178E mutants, respectively; ( B ) statistical analysis of hIL-6 mutants compared with hIL-6 WT in term of STAT3 phosphorylation levels in HepG2 cells, based on three independent experiments. Data is presented as mean ± SD, n = 3. **p < 0.01; and ***p < 0.001.

Article Snippet: Subsequently, the cells were washed and stained for 30 min at room temperature in the dark with anti-pSTAT3 (pY705)-PE and an anti-mouse immunoglobulin G isotype-matched control (BD, Cat, 558595).

Techniques: Activation Assay, Negative Control, Western Blot, Expressing

Journal: Scientific Reports

Article Title: Unveiling novel insights into human IL-6 − IL-6R interaction sites through 3D computer-guided docking and systematic site mutagenesis

doi: 10.1038/s41598-024-69429-w

Figure Lengend Snippet: Activate of phosphorylation of STAT3 in Leukocytes by IL-6 mutants. Human leukocytes were preincubated with 50 ng/mL IL-6 mutants for 15 min, followed by permeabilized and stained. Untreated cells were used as the negative control (NC). Phosphorylation of STAT3 was assessed using flow cytometry with PE labeled anti-pSTAT3 (pY705) monoclonal antibody. ( A – C ) Gating strategy and representative histogram graph of P-STAT3 + cells.; ( D ) statistical analysis of percentage of P-STAT3 + cells upon stimulation with hIL-6 WT or R167A, E171A and R178E mutants, based on three independent experiments. The data is presented as mean ± SD, n = 3. *p < 0.05; **p < 0.01.

Article Snippet: Subsequently, the cells were washed and stained for 30 min at room temperature in the dark with anti-pSTAT3 (pY705)-PE and an anti-mouse immunoglobulin G isotype-matched control (BD, Cat, 558595).

Techniques: Staining, Negative Control, Flow Cytometry, Labeling

Journal: bioRxiv

Article Title: LDL-Binding IL-10 Reduces Vascular Inflammation in Atherosclerotic Mice

doi: 10.1101/2024.03.04.582839

Figure Lengend Snippet: Fusing IL-10 to an LDL-binding antibody fragment enables lipoprotein binding while maintaining signaling and anti-inflammatory properties. a) Conceptual schematic of plaque-targeted IL-10. b) SDS PAGE gel showing various Fab-IL-10 constructs. N, non-reducing conditions. R, reducing conditions. c-e) Binding affinity of Fab-IL-10 to LDL measured using surface plasmon resonance. KD, dissociation constant. Dashed lines represented calculated fit. f) Activity of IL-10 and Fab-IL-10 by phosphorylation of STAT3 (pSTAT3), measured by flow cytometry of RAW 264.7 cells incubated with the indicated concentrations of IL-10 (n = 3). g) Log(EC50) calculated from fitted curves in (f), shown with 95% confidence intervals. h) TNFα secretion of LPS-stimulated RAW 264.7 cells incubated with WT IL-10 or Fab-IL-10 (n = 5). Experiments were performed twice with similar results. Data represent mean +/-standard deviation (f) or mean + standard deviation (h). Statistics performed by one-way ANOVA with Dunnett’s post-test compared to media.

Article Snippet: Cells were stained with PE-conjugated antibodies against pSTAT3 (pY705, clone 4/P-STAT3, BD Biosciences) at a 1:50 dilution for 1 hr at room temperature in the dark.

Techniques: Binding Assay, SDS Page, Construct, SPR Assay, Activity Assay, Flow Cytometry, Incubation, Standard Deviation

Journal: Cell

Article Title: Human MCTS1-dependent translation of JAK2 is essential for IFN-γ immunity to mycobacteria

doi: 10.1016/j.cell.2023.09.024

Figure Lengend Snippet: Key resources table

Article Snippet: Human pSTAT3-PE (pY705) , BD , Cat# 612569, RRID:AB_399860.

Techniques: Virus, Recombinant, Staining, Selection, Polymerase Chain Reaction, Plasmid Preparation, Reporter Assay, Sequencing, Mutagenesis, Variant Assay, Clone Assay, Knock-Out, Software, Next-Generation Sequencing