biotinylated anti mouse tlr9  (Hycult Biotech)

Journal: Malaria Journal

Article Title: Pre-existing Schistosoma japonicum infection alters the immune response to Plasmodium berghei infection in C57BL/6 mice

doi: 10.1186/1475-2875-12-322

Figure Lengend Snippet: Mice in Plasmodium berghei ANKA-mono-infection group and Schistosoma japonicum-P. berghei ANKA-co-infection group were compared at different time points according to data obtained by flow cytometry. Proportion of splenic DCs subpopulation, CD11c + CD11b + and CD11c + CD45R⁄ B220 + , were analysed in A and B , with absolute number of these DCs present in C and D . Proportion of splenic CD11c + DCs expressing TLR4 , TLR9, MHC class II, and CD86 of the two groups were compared in E , F , G , H , respectively. MFI of TLR4, TLR9, MHC class II, and CD86, which were expressed on CD11c + CD11b + DCs and CD11c + CD45R⁄ B220 + DCs were present in I and J . At each time point, at least three mice per group were sacrificed. Bars represent the mean values ± SD. Student t test by comparisons between the two groups was performed with * indicating P <0.05 and # indicating P <0.01.

Article Snippet: After fixation and permeabilization using staining buffer reagents as instructed by the manufacturer, cells were incubated with biotinylated anti-mouse TLR9 (clone 5G5, Hycult Biotechnology (HBT), Uden, The Netherlands) followed by PE-conjugated streptavidin (Biolegend, San Diego, CA, USA).

Techniques: Infection, Flow Cytometry, Expressing

Journal: Journal of Neuroinflammation

Article Title: TLR2 and TLR9 modulate enteric nervous system inflammatory responses to lipopolysaccharide

doi: 10.1186/s12974-016-0653-0

Figure Lengend Snippet: TLR2/4/9 are expressed in enteric neurons of adult rat colon tissue. TLR localisation in whole-mount preparations of the SMP and the LMMP. B-tubulin III was used instead of HuC/D as neuronal marker for colocalisation with TLR9 due to antibody detection incompatibilities. Scale bars : 25 μm

Article Snippet: Primary antibodies used in immunofluorescence were rabbit monoclonal anti-TLR2 (1:500; Abcam, Cambridge, UK), rabbit polyclonal anti-TLR4 (1:100; Novus Biologicals, Cambridge, UK), mouse monoclonal anti-TLR9 (1:100; Novus Biologicals), chicken polyclonal anti-GFAP (1:500; Antibodies-online, Aachen, Germany), mouse monoclonal anti- S100β (1:1000; Abcam), mouse monoclonal anti-HuC/D (1:200; Life Technologies), chicken polyclonal anti-β-tubulin III (1:500; Abcam), rabbit monoclonal anti-NF-kB p65 (1:50; Cell Signaling Technology, Danvers, USA), goat polyclonal anti-IL-6 (1:250; Santa Cruz Biotechnology), mouse monoclonal anti-MCP-1 (1:100; Novus Biologicals) and rabbit polyclonal anti- ionized calcium-binding adapter molecule (IBA)-1 (1:500; Wako Chemicals).

Techniques: Marker

Journal: Journal of Neuroinflammation

Article Title: TLR2 and TLR9 modulate enteric nervous system inflammatory responses to lipopolysaccharide

doi: 10.1186/s12974-016-0653-0

Figure Lengend Snippet: Embryonic ENS culture neurons express TLR2/4/9. a Agarose gel showing specific products of real-time PCR for the assayed genes in ENS culture (PC); water was used as a no-template control (H 2 O) and rat colon cDNA as positive control (+C). b TLR relative expression in ENS culture in basal conditions ( n = 8). c Localisation of TLR in ENS culture ganglia. B-tubulin III was used instead of HuC/D as neuronal marker for colocalisation with TLR9 due to antibody detection incompatibilities. White arrows point to neuronal somas. Scale bars : 25 μm

Article Snippet: Primary antibodies used in immunofluorescence were rabbit monoclonal anti-TLR2 (1:500; Abcam, Cambridge, UK), rabbit polyclonal anti-TLR4 (1:100; Novus Biologicals, Cambridge, UK), mouse monoclonal anti-TLR9 (1:100; Novus Biologicals), chicken polyclonal anti-GFAP (1:500; Antibodies-online, Aachen, Germany), mouse monoclonal anti- S100β (1:1000; Abcam), mouse monoclonal anti-HuC/D (1:200; Life Technologies), chicken polyclonal anti-β-tubulin III (1:500; Abcam), rabbit monoclonal anti-NF-kB p65 (1:50; Cell Signaling Technology, Danvers, USA), goat polyclonal anti-IL-6 (1:250; Santa Cruz Biotechnology), mouse monoclonal anti-MCP-1 (1:100; Novus Biologicals) and rabbit polyclonal anti- ionized calcium-binding adapter molecule (IBA)-1 (1:500; Wako Chemicals).

Techniques: Agarose Gel Electrophoresis, Real-time Polymerase Chain Reaction, Control, Positive Control, Expressing, Marker

Journal: Journal of Neuroinflammation

Article Title: TLR2 and TLR9 modulate enteric nervous system inflammatory responses to lipopolysaccharide

doi: 10.1186/s12974-016-0653-0

Figure Lengend Snippet: TLR activation in ENS culture is associated to TLR2 up-regulation. TLR mRNA levels were assessed in rat ENS culture stimulated for 8 h with the indicated MAMPs. a Pam2CSK4 ( n = 5–10; TLR2 ligand vs. control, *** P < 0.001). b LPS ( n = 5–10; TLR2 ligand vs. control, *** P < 0.001; TLR4 and TLR9 ligand vs. control, * P < 0.05). c ODN 1826 ( n = 5–10; TLR2 ligand vs. control, ** P < 0.01; TLR9 ligand vs. control, * P < 0.05)

Article Snippet: Primary antibodies used in immunofluorescence were rabbit monoclonal anti-TLR2 (1:500; Abcam, Cambridge, UK), rabbit polyclonal anti-TLR4 (1:100; Novus Biologicals, Cambridge, UK), mouse monoclonal anti-TLR9 (1:100; Novus Biologicals), chicken polyclonal anti-GFAP (1:500; Antibodies-online, Aachen, Germany), mouse monoclonal anti- S100β (1:1000; Abcam), mouse monoclonal anti-HuC/D (1:200; Life Technologies), chicken polyclonal anti-β-tubulin III (1:500; Abcam), rabbit monoclonal anti-NF-kB p65 (1:50; Cell Signaling Technology, Danvers, USA), goat polyclonal anti-IL-6 (1:250; Santa Cruz Biotechnology), mouse monoclonal anti-MCP-1 (1:100; Novus Biologicals) and rabbit polyclonal anti- ionized calcium-binding adapter molecule (IBA)-1 (1:500; Wako Chemicals).

Techniques: Activation Assay, Control

Journal: Arthritis Research & Therapy

Article Title: Overexpression of TLR2 and TLR9 on monocyte subsets of active rheumatoid arthritis patients contributes to enhance responsiveness to TLR agonists

doi: 10.1186/s13075-015-0901-1

Figure Lengend Snippet: Increased level of toll-like receptor 9 ( TLR9 ) on circulating and synovial monocyte subsets of rheumatoid arthritis ( RA ) patients. a Flow cytometry analysis of TLR9 expression on blood monocyte subsets from healthy controls and RA patients. TLR9 expression for synovial monocyte subsets is also shown for RA patients. TLR9 expression appears in green (P1), blue (P2) or purple (P3). Gray histograms represent internal negative controls. Data are cell populations of representative donors. b TLR9 expression on blood monocyte subsets from healthy controls and RA patients are presented as mean fluorescence intensity ( MFI ) (mean ± standard error of the mean (SEM)). Black bars healthy donors, gray bars RA patients. c TLR9 expression in the monocyte subsets of RA patient synovial fluids is presented as MFI (mean ± SEM). Striped bars RA patient synovial fluids: * p ≤0.05

Article Snippet: Enriched monocyte subsets (identified as detailed above) and neutrophils were washed twice in Hanks Balanced Salt Sodium (HBSS; Wisent) and stained with TLR2-fluorescein isothiocyanate (FITC) (TL2.1) or TLR9-FITC (5G5) antibodies (Hycult Biotech, Uden, The Netherlands).

Techniques: Flow Cytometry, Expressing, Fluorescence

Journal: Arthritis Research & Therapy

Article Title: Overexpression of TLR2 and TLR9 on monocyte subsets of active rheumatoid arthritis patients contributes to enhance responsiveness to TLR agonists

doi: 10.1186/s13075-015-0901-1

Figure Lengend Snippet: Expression of toll-like receptor 2 ( TLR2 ) and TLR 9 in neutrophils of rheumatoid arthritis ( RA ) patients. a Flow cytometry analysis of TLR2 and TLR9 expression in neutrophils isolated from blood of healthy controls and RA patients and synovial fluids of RA patients. TLR2 and TLR9 expression is represented by red histograms whereas negative control is represented by gray histograms . Data are neutrophils of representative donors. b TLR2 and TLR9 expression on blood neutrophils from healthy controls and RA patients are presented as mean fluorescence intensity ( MFI ) (mean ± standard error of the mean (SEM)). Black bars healthy donors, gray bars RA patients . c TLR2 and TLR9 expression on synovial fluid neutrophils from RA patients are presented as MFI (mean ± SEM). Striped bars RA patient synovial fluids

Article Snippet: Enriched monocyte subsets (identified as detailed above) and neutrophils were washed twice in Hanks Balanced Salt Sodium (HBSS; Wisent) and stained with TLR2-fluorescein isothiocyanate (FITC) (TL2.1) or TLR9-FITC (5G5) antibodies (Hycult Biotech, Uden, The Netherlands).

Techniques: Expressing, Flow Cytometry, Isolation, Negative Control, Fluorescence

Journal: Arthritis Research & Therapy

Article Title: Overexpression of TLR2 and TLR9 on monocyte subsets of active rheumatoid arthritis patients contributes to enhance responsiveness to TLR agonists

doi: 10.1186/s13075-015-0901-1

Figure Lengend Snippet: Intracellular cytokine expression in monocyte subsets from rheumatoid arthritis (RA) patients following toll-like receptor 9 (TLR9) stimulation. Mononuclear cells from RA patients and healthy donors were enriched and stimulated with CpG-2006 ( a ) and Epstein–Barr virus ( EBV ) DNA (TLR9 ligands) ( b ) in vitro. Gating strategy is described in Fig. . Intracellular cytokine level for IL-6, IL-1β, TNFα and monocyte chemoattractant protein-1 ( MCP-1 ) are shown for all three subsets of monocytes. Gray non-stimulated samples, Blue healthy donors, red RA patients. Representative cytometry histograms are representative of ten donors

Article Snippet: Enriched monocyte subsets (identified as detailed above) and neutrophils were washed twice in Hanks Balanced Salt Sodium (HBSS; Wisent) and stained with TLR2-fluorescein isothiocyanate (FITC) (TL2.1) or TLR9-FITC (5G5) antibodies (Hycult Biotech, Uden, The Netherlands).

Techniques: Expressing, In Vitro, Cytometry