rabbit polyclonal anti irak4  (Cell Signaling Technology Inc)

Journal: Cell Reports Medicine

Article Title: Osteoclast-derived apoptotic bodies inhibit naive CD8 + T cell activation via Siglec15, promoting breast cancer secondary metastasis

doi: 10.1016/j.xcrm.2023.101165

Figure Lengend Snippet: Siglec15 inhibits naive CD8 + T cell activation via blocking TLR2 co-stimulatory signaling pathways (A) Representative flow cytometry results of CD8 + T cell (stained with CFSE) exposed to different treatments. (B) Quantification of CD8 + T cell division and immune secretion (IFN-γ, TNF-α, IL-2; per well of 96-well plate), n = 3. (C) Western blot result showed expression of IRAK4, TRAF6, and TAK1 in CD8 + T cell lysate after different treatments, and quantification of the relative expression of TRAF6 in CD8 + T cells under different treatments, n = 3. (D) Representative IHC for IRAK4, TRAF6, and TAK1 in bone metastases sections from three groups of mice. Scale bar: 200 μm. (E) Semi-quantitative analysis of IRAK4, TRAF6, and TAK1 IHC staining images, n = 5. The data in each panel represent the means ± SD and were analyzed by Student’s two-tailed unpaired t test, ∗p < 0.05, ∗∗p < 0.01.

Article Snippet: Rabbit anti-IRAK4 , Bioss , Cat#bs-2440R;RRID: AB_11067218.

Techniques: Activation Assay, Blocking Assay, Flow Cytometry, Staining, Western Blot, Expressing, Immunohistochemistry, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Osteoclast-derived apoptotic bodies inhibit naive CD8 + T cell activation via Siglec15, promoting breast cancer secondary metastasis

doi: 10.1016/j.xcrm.2023.101165

Figure Lengend Snippet:

Article Snippet: Rabbit anti-IRAK4 , Bioss , Cat#bs-2440R;RRID: AB_11067218.

Techniques: Virus, Recombinant, Plasmid Preparation, Staining, Cell Isolation, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Purification, Bicinchoninic Acid Protein Assay, Isolation, Software

Journal: Journal of Traditional Chinese Medicine

Article Title: Efficacy of Sishen Wan (四神丸) on dinitrobenzene sulfonic acid-induced ulcerative colitis and its effect on toll-like receptor 2/interleukin-1 receptor-associated kinase-4/nuclear factor-κB signal pathway

doi: 10.19852/j.cnki.jtcm.20220608.001

Figure Lengend Snippet: primer sequence

Article Snippet: Dinitrobenzene sulfonic acid (DNBS) (Sigma-Aldrich, St. Louis, MO, USA); hematoxylin-eosin (HE) staining Kit (Solarbio Science & Technology Co., Ltd., Beijing, China); terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) apoptosis detection kit (Yeasen Biotechnology Co., Ltd., Shanghai, China); immunohistochemical staining kit (ZSGB-Bio Co., Ltd., Beijing, China); rat tumor necrosis factor-α (TNF-α) enzyme linked immunosorbent assay (ELISA) kit (Enzyme-linked Biotechnology Co., Ltd., Shanghai, China); rat interleukin-1β (IL-1β) ELISA kit (Enzyme-linked Biotechnology Co., Ltd., Shanghai, China); radio immunoprecipitation assay (RIPA) lysate (Solarbio Science & Technology Co., Ltd., Beijing, China); nuclear protein extraction kit (Solarbio Science & Technology Co., Ltd., Beijing, China); total RNA extraction kit (Takara Biomedical Technology Co., Ltd., Beijing, China); PrimeScript™ RT reagent kit with gDNA rraser (Perfect Real Time) (Takara Biomedical Technology Co., Ltd., Beijing, China); TB Green® Premix Ex Taq™ (Tli RNaseH Plus) (Takara Biomedical Technology Co., Ltd., Beijing, China); Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis (SDS-PAGE) Gel kit Solarbio Science & Technology Co., Ltd., Beijing, China); rabbit anti-TLR2 polyclonal antibody (Huabio, Hangzhou, China), rabbit anti-IRAK4 polyclonal antibody (Bioss, Beijing, China), rabbit anti-NF-κB p65 polyclonal antibody (CST, Danforth, MA, USA), rabbit anti phospho-NF-κB p65 (Ser468) antibody (Bioss, Beijing, China), rabbit anti-MyD88 polyclonal antibody (Abcam Cambridge, UK), rabbit anti β-actin antibody (Abcam Cambridge, UK).

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Euphorbia Factor L2 ameliorates the Progression of K/BxN Serum-Induced Arthritis by Blocking TLR7 Mediated IRAK4/IKKβ/IRF5 and NF-kB Signaling Pathways

doi: 10.3389/fphar.2021.773592

Figure Lengend Snippet: EFL2 reduced IL-1β and IL-6 production via robustly inhibiting TLR7-IKKβ-IRF5 signalling pathway (A-C) Western blot bands (A) and quantification of p -IRAK4, IRAK4 (B) , p -IKKβ and IKKβ expression (C) in RAW264.7 cells (D-F) Western blot bands (D) and effects of EFL2 on R837-induced IRF5 phosphorylation (E) and IRF5 expression in cytoplasm and nucleus (F) (G) IRF5 localization determined by immunofluorescence staining (Magnification: ×40) (H) IRF5 expression in cytoplasm and nucleus after IRF5 overexpression (I) Quantification of IRF5 expression after transfection (J, K) The levels of IL-1β (J) and IL-6 (K) produced by RAW264.7 cells before and after IRF5 overexpression (n = 3 for each group in an independent experiment). Data represent the mean values ±SEM of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 were considered as significant.

Article Snippet: The antibodies used in the experiments were as follows: rabbit anti-mouse IKKα/β polyclonal antibody (1:1,000, Abways, China), rabbit anti-mouse IκBα polyclonal antibody (1:1,000, Abgent, China), rabbit anti-mouse p65 polyclonal antibody (1:1,000, Abgent, China), rabbit anti-mouse phosphorylated IKKα (Ser176)/β (Ser177) (1:1,000, Abways, China), rabbit anti-mouse phosphorylated IκBα (1:1,000, Abcam, United States), rabbit anti-mouse phosphorylated p65 (1:1,000, Abcam, United States), rabbit anti-mouse IRAK4 polyclonal antibody (1:1,000, Proteintech, United States), rabbit anti-mouse phosphorylated IRAK4 (Thr345/Ser346) (1:1,000, Affinity, United States), rabbit anti-mouse IKKβ polyclonal antibody (1:1,000, Abbkine, United States), rabbit anti-mouse phosphorylated IKKβ (Ser177) (1:1,000, Abways, China), rabbit anti-mouse IRF5 polyclonal antibody (1:1,000, Immunoway, United States), rabbit anti-mouse phosphorylated IRF5 (Ser437) (1:1,000, Affinity, United States), anti-mouse actin monoclonal antibody (1:20,000, Transgen, China) and rabbit anti-mouse LaminB monoclonal antibody (1:5,000, Abways, China).

Techniques: Western Blot, Expressing, Phospho-proteomics, Immunofluorescence, Staining, Over Expression, Transfection, Produced