mouse anti tlr 4  (Santa Cruz Biotechnology)

Journal: International Journal of Molecular Sciences

Article Title: BDMC33, A Curcumin Derivative Suppresses Inflammatory Responses in Macrophage-Like Cellular System: Role of Inhibition in NF-κB and MAPK Signaling Pathways

doi: 10.3390/ijms13032985

Figure Lengend Snippet: Effects of BDMC33 on IFN-γ/LPS-treated surface expression of TLR-4 and CD-14 accessory molecules in RAW 264.7 macrophages. Cells were stimulated with IFN-γ/LPS and treated with increasing concentrations of BDMC33 for 16 h. The cells were then collected and stained with FITC-conjugated anti-mouse CD-14 and PerCP-conjugated anti-mouse TLR-4 and acquired on FACSCalibur flow cytometer. The expression level was normalized against a IFN-γ/LPS-treated group. C; Basal level of CD-14 or TLR-4 without IFN-γ/LPS treatment. The results are expressed in mean ± S.E.M. of three independent experiments. * P < 0.05, significantly different from IFN-γ/LPS-treated control group (second column).

Article Snippet: Rabbit polyclonal against human p65, ERK1/2, pERK1/2, p38, pp38, I-κB, pI-κB, HRP conjugated anti-β-actin, fluorescein isothiocynate (FIT-C) conjugated anti-mouse TLR-4 and Peridinin chlorophyll protein complex (PerCP) conjugated anti-mouse CD-14 from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Staining, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: BDMC33, A Curcumin Derivative Suppresses Inflammatory Responses in Macrophage-Like Cellular System: Role of Inhibition in NF-κB and MAPK Signaling Pathways

doi: 10.3390/ijms13032985

Figure Lengend Snippet: A schematic diagram shows the mechanisms underlying the inhibitory action of BDMC33. TLR-4 stimulation by LPS activates an intracellular signaling cascade that involves the recruitment of MyD88 (myeloid differentiation primary response gene-88) and results in the phosphorylation of TRAF-6 (tumor-necrosis-factor-receptor-associated factor 6), subsequently activate NF-κB and MAPK pathways. The red blunt lines indicate the inhibition by BDMC33.

Article Snippet: Rabbit polyclonal against human p65, ERK1/2, pERK1/2, p38, pp38, I-κB, pI-κB, HRP conjugated anti-β-actin, fluorescein isothiocynate (FIT-C) conjugated anti-mouse TLR-4 and Peridinin chlorophyll protein complex (PerCP) conjugated anti-mouse CD-14 from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Inhibition

Journal: Journal of Immunology Research

Article Title: Characterization of Toll-Like Receptor-4 (TLR-4) in the Spleen and Thymus of Swiss Albino Mice and Its Modulation in Experimental Endotoxemia

doi: 10.1155/2015/137981

Figure Lengend Snippet: mRNA expression of TLR-4 in the spleen and thymus: the results in this figure (a) represents the differential transcription of tlr-4 gene on LPS administration in thymus and spleen of Swiss albino mice from triplicate experiments; lane 1 represents TLR-4 mRNA in thymus from control mice (CT), lane 2 thymus from LPS treated mice at 6 hours after treatment (LT6), lane 3 thymus from LPS treated mice at 12 hours after treatment (LT12), lane 4 thymus from LPS treated mice at 24 hours after treatment (LT24), lane 5 represents spleen from control mice (CS), lane 6 spleen from LPS treated mice at 6 hours after treatment (LS6), lane 7 spleen from LPS treated mice at 12 hours after treatment (LS12), and lane 8 spleen from LPS treated mice at 24 hours post treatment (LS24). (b) Is the diagrammatic representation of the fold difference in their expression compare to control and among different time periods of treatment. Molecular weight marker (100 bp). The following symbols indicates the significant change in the expression of TLR-4 between: * control versus LPS treated tissue, # LPS treatment for 6 hours versus LPS treatment for 12 and 24 hours, % LPS treatment for 12 hours versus LPS treatment 24 hours. Significant change ( P < 0.05).

Article Snippet: The membranes were blocked with 5% BSA in TBST for 3 hour at room temperature, washed and incubated with primary anti-mice TLR-4 Abs in 1/1000 dilution (cat no- ab13867, Abcam, UK, synthetic peptide corresponding to amino acid 39–56 of mouse TLR-4) overnight at 4°C.

Techniques: Expressing, Molecular Weight, Marker

Journal: Journal of Immunology Research

Article Title: Characterization of Toll-Like Receptor-4 (TLR-4) in the Spleen and Thymus of Swiss Albino Mice and Its Modulation in Experimental Endotoxemia

doi: 10.1155/2015/137981

Figure Lengend Snippet: Expression of TLR-4 receptor protein in the spleen and thymus. The results in Figures (a) and (c) represent the differential translation of the TLR-4 mRNA to the protein in the spleen and thymus, respectively, in mice with experimental endotoxemia from triplicate experiments. Lane 1 (Con) represents TLR-4 in tissues from control mice, lane 2 (LPS3H) from LPS treated mice at 3 hours after treatment, lane 3 (LPS6H) from LPS treated mice at 6 hours after treatment, lane 4 (LPS12H) from LPS treated mice at 12 hours after treatment, lane 5 (LPS24H) from LPS treated mice at 24 hours after treatment, lane 6 (LPS48H) from LPS treated mice at 48 hours after treatment, and lane 7 (LPS72H) from LPS treated mice at 72 hours after treatment. Results in (b) and (d) is the diagrammatic representation of the fold difference in their expression after LPS treatment compared to control and among different time periods of treatment as seen in (a) and (c), respectively. The approximate band size is about 95 KD. The following symbols indicates the significant change in the expression of TLR-4 between: * control versus LPS treated tissues, # LPS treatment for 3 hours versus LPS treatment for 6, 12, 24, 48, and 72 hours, % LPS treatment for 6 hours versus LPS treatment for 12, 24, 48, and 72 hours, $ LPS treatment for 12 hours versus LPS treatment for 24, 48, and 72 hours. Significant change ( P < 0.05).

Article Snippet: The membranes were blocked with 5% BSA in TBST for 3 hour at room temperature, washed and incubated with primary anti-mice TLR-4 Abs in 1/1000 dilution (cat no- ab13867, Abcam, UK, synthetic peptide corresponding to amino acid 39–56 of mouse TLR-4) overnight at 4°C.

Techniques: Expressing

Journal: Journal of Immunology Research

Article Title: Characterization of Toll-Like Receptor-4 (TLR-4) in the Spleen and Thymus of Swiss Albino Mice and Its Modulation in Experimental Endotoxemia

doi: 10.1155/2015/137981

Figure Lengend Snippet: Expression of TLR-4 receptor in purified splenic macrophages and lymphocytes after in vitro stimulation with LPS. The results in (a) and (c) show the differential translation of the TLR-4 mRNA to the protein in the splenic macrophages and lymphocytes, respectively, after in vitro stimulation with LPS from a set of triplicate experiments. Lane 1 (M φ Con) represents TLR-4 on untreated macrophages, lane 2 (M φ + L60 m) represents TLR-4 expression on the macrophages treated with LPS for 60 minutes, lane 3 (M φ + L90 m) represents TLR-4 expression on macrophages treated with LPS for 90 minutes, and lane 4 (M φ + 120 m) represents TLR-4 expression on macrophages treated with LPS for 120 minutes. Lane 1 (Lym Con) represents TLR-4 on untreated lymphocyte, lane 2 (Lym + L60 m) represents TLR-4 expression on the lymphocyte treated with LPS for 60 minutes, lane 3 (Lym + L90 m) represents TLR-4 expression on lymphocyte treated with LPS for 90 minutes, and lane 4 (Lym + L120 m) represents TLR-4 expression on lymphocyte treated with LPS for 120 minutes. The following symbols represent the significant fold change in TLR-4 expression between: * control versus LPS treated cells, # LPS treatment for 60 min versus LPS treatment for 90 and 120 min, % LPS treatment for 90 min versus LPS treatment for 120 min. Significant change ( P < 0.05).

Article Snippet: The membranes were blocked with 5% BSA in TBST for 3 hour at room temperature, washed and incubated with primary anti-mice TLR-4 Abs in 1/1000 dilution (cat no- ab13867, Abcam, UK, synthetic peptide corresponding to amino acid 39–56 of mouse TLR-4) overnight at 4°C.

Techniques: Expressing, Purification, In Vitro

Journal: Mediators of Inflammation

Article Title: Extracellular Cardiolipin Modulates Select Immune Functions of Astrocytes in Toll-Like Receptor (TLR) 4-Dependent Manner

doi: 10.1155/2022/9946439

Figure Lengend Snippet: The effect of CL on the expression of TLR 4 by unstimulated (a) and LPS-stimulated (b) human U118 MG astrocytic cells. CL (20 μ g/ml) or its vehicle solution was added to U118 MG astrocytic cell cultures 15 min before exposure to LPS (400 ng/ml) or its vehicle solution. After 48 h, TLR 4 protein levels were measured by immunostaining. The resulting chemiluminescence signal was normalized to protein concentration in each well. Data (means ± SEM) are presented as a ratio of normalized chemiluminescence signal to that obtained from untreated control cells. ∗ P < 0.05 according to the paired Student's t -test.

Article Snippet: Anti-mouse postsynaptic density protein (PSD)-95 antibody (#MA1-045), mouse anti-TLR 4 antibodies (#5015348; clone HTA125), and all other reagents were purchased from Fisher Scientific (Ottawa, ON, Canada).

Techniques: Expressing, Immunostaining, Protein Concentration

Journal: Mediators of Inflammation

Article Title: Extracellular Cardiolipin Modulates Select Immune Functions of Astrocytes in Toll-Like Receptor (TLR) 4-Dependent Manner

doi: 10.1155/2022/9946439

Figure Lengend Snippet: The effect of TLR 4-specific antagonist TAK-242 on CL-induced secretion of MCP-1 by human U118 MG astrocytic cells and their viability in serum-containing (a, c) and serum-free (b, d) media. TAK-242 (10 μ M) was added to U118 MG astrocytic cells 30 min before their exposure to CL. After 48 h, the concentration of MCP-1 in cell supernatants was measured by an ELISA, and the viability of U118 MG astrocytic cells was measured by the MTT assay. Data are presented as means ± SEM. Cell viability in (c, d) is normalized to values obtained from U118 MG cultures not exposed to CL, TAK-242, or their vehicle solutions. ∗ P < 0.05 and ∗∗ P < 0.01 according to Tukey's post-hoc test. P and F values for the one-way randomized blocks ANOVA are shown. The detection limit of the ELISA is presented as the dotted line.

Article Snippet: Anti-mouse postsynaptic density protein (PSD)-95 antibody (#MA1-045), mouse anti-TLR 4 antibodies (#5015348; clone HTA125), and all other reagents were purchased from Fisher Scientific (Ottawa, ON, Canada).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, MTT Assay

Journal: PLoS ONE

Article Title: The Role of Lipid Raft Aggregation in the Infection of Type II Pneumocytes by Mycobacterium tuberculosis

doi: 10.1371/journal.pone.0045028

Figure Lengend Snippet: A549 cells were infected with live Mtb strains CDC1551, HN878 or H37Rv at MOI 100 or treated with total lipids (TL), culture filtrate proteins (CFP) and gamma-irradiated cells for 24 hr, as described. Colocalization of anti-TLR-2 and anti-TLR-4 antibodies with LR aggregates was analyzed from confocal images using Manders coefficient values. Infections with CDC1551, HN878 and H37Rv produced significantly less TLR-2 and TLR-4 colocalization compared to positive controls (*p-value <0.001) (A). There were no significant differences between TL-treated cells and DMSO controls at this time point (B) (TLR-4, p-value = 0.532 to 0.983; TLR-2, p-value = 0.775 to 1.0). TLR-2 and TLR-4 colocalization for controls and CFP-treated cells was comparable (C). At 24 hr post-TL (B) and –CFP (C) treatment, colocalization of receptors was significantly decreased compared to positive controls (*p-value <0.001). These findings also held true for epithelial cells infected with gamma-irradiated bacteria from all three strains (*p-value <0.001) (D). Images were collected at 63x magnification and analyzed using ImageJ JACoP plugin. Infections were performed in duplicate and repeated three times.

Article Snippet: Cells were incubated with either mouse monoclonal anti-TLR-4 (Santa Cruz) or mouse monoclonal anti-TLR-2 (Abcam) antibodies at 1∶200 dilution for 1 hr at RT.

Techniques: Infection, Irradiation, Produced