rabbit polyclonal antibodies against tlr4  (Santa Cruz Biotechnology)

Journal: Pathogens

Article Title: The Immunological Changes in the Skin of BALC/c Mice with Disseminated Acanthamoebiasis

doi: 10.3390/pathogens12050631

Figure Lengend Snippet: The TLR2 gene expression in the skin of control and amoeba-infected mice at 8, 16 and 24 dpi, (A, amoeba-infected host with normal immunity; AS, amoeba-infected host with reduced immunity induced by MPS, methylprednisolone sodium succinate; C, control host with normal immunity; CS, control host with reduced immunity induced by MPS). In the graph arithmetic mean ± standard deviation was shown; p < 0.05.

Article Snippet: Immunohistochemistry was performed using specific primary rabbit polyclonal antibodies against TLR2 and TLR4 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; cat. no. sc-10739 and sc-30002, respectively) in a final 1:500 dilution, after checking and standardized other recommended by manufacturer concentrations from 1:50 to 1:500.

Techniques: Expressing, Infection, Standard Deviation

Journal: Pathogens

Article Title: The Immunological Changes in the Skin of BALC/c Mice with Disseminated Acanthamoebiasis

doi: 10.3390/pathogens12050631

Figure Lengend Snippet: Representative microphotography that shows immunoexpression of TLR2 ( A – H ) and TLR4 ( I – P ) in skin of immunocompetent and immunosuppressed uninfected mice (0 dpi) and at 8, 16, and 24 days after Acanthamoeba spp. infection (dpi). Immunohistochemical reaction with diaminobenzidine (DAB) as a chromogen. Objective magnification 40×. Red arrows mark the area of connective tissue of dermis where immunoexpression of Toll-like receptors 2 (TLR2) and 4 (TLR4) was visible as brown pigmentation; white arrows mark the keratinocytes of epidermis where immunoexpression of TLR2 was visible as brown pigmentation. Abbreviation: C—uninfected immunocompetent mice (n = 5), CS—uninfected immunosuppressed mice (n = 5), A—immunocompetent Acanthamoeba spp. infected mice (n = 5), AS—immunosuppressed Acanthamoeba spp. infected mice (n = 5).

Article Snippet: Immunohistochemistry was performed using specific primary rabbit polyclonal antibodies against TLR2 and TLR4 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; cat. no. sc-10739 and sc-30002, respectively) in a final 1:500 dilution, after checking and standardized other recommended by manufacturer concentrations from 1:50 to 1:500.

Techniques: Infection, Immunohistochemical staining

Journal: PLoS ONE

Article Title: The Toll-Like Receptor 4 Polymorphism Asp299Gly but Not Thr399Ile Influences TLR4 Signaling and Function

doi: 10.1371/journal.pone.0093550

Figure Lengend Snippet: Lentiviral vectors and cell lines stably expressing these vectors.

Article Snippet: The following antibodies were used: mouse anti-human TLR4-APC antibody, mouse anti-human TLR4 purified antibody, rat IgG2a K isotype control APC, mouse IgG1 isotype control Alexa Fluor647 and mouse IgG2b isotype control Alexa Fluor647 (eBioscience); rat anti-mouse CD16/CD32, anti-NF-κB p65 antibody (BD Biosciences); PerCP anti-human IL-8 and PerCP mouse Ig G2a isotype control (Biolegend); horseradish peroxidase-conjugated goat anti-rabbit IgG (Invitrogen/LifeTechnologies); rabbit polyclonal antibody against human TLR4 (Santa Cruz Biotechnology, Inc); and rabbit monoclonal antibody against human β-actin (Cell Signaling).

Techniques: Stable Transfection, Expressing

Journal: PLoS ONE

Article Title: The Toll-Like Receptor 4 Polymorphism Asp299Gly but Not Thr399Ile Influences TLR4 Signaling and Function

doi: 10.1371/journal.pone.0093550

Figure Lengend Snippet: Each cell line was examined for total ( A, D ), cell surface ( B, E ) and intracellular ( C, F ) TLR4 expression by flow cytometry using the mouse anti-human TLR4-APC antibody. Percentage of total, cell surface and intracellular TLR4 positive cells for cell line groups are shown in panels ( A – C ). Displayed in panels ( D – F ) are mean fluorescent intensities (MFI) of TLR4 positive cells in each genotype group. All data are presented as mean ± SEM of three independent experiments. Statistical comparisons were performed using One-way ANOVA and Newman-Keuls post-hoc test among the TLR4 genotype groups (n = 3) except the Asp299Gly/Thr399Ile cell group (n = 2).

Article Snippet: The following antibodies were used: mouse anti-human TLR4-APC antibody, mouse anti-human TLR4 purified antibody, rat IgG2a K isotype control APC, mouse IgG1 isotype control Alexa Fluor647 and mouse IgG2b isotype control Alexa Fluor647 (eBioscience); rat anti-mouse CD16/CD32, anti-NF-κB p65 antibody (BD Biosciences); PerCP anti-human IL-8 and PerCP mouse Ig G2a isotype control (Biolegend); horseradish peroxidase-conjugated goat anti-rabbit IgG (Invitrogen/LifeTechnologies); rabbit polyclonal antibody against human TLR4 (Santa Cruz Biotechnology, Inc); and rabbit monoclonal antibody against human β-actin (Cell Signaling).

Techniques: Expressing, Flow Cytometry

Journal: PLoS ONE

Article Title: The Toll-Like Receptor 4 Polymorphism Asp299Gly but Not Thr399Ile Influences TLR4 Signaling and Function

doi: 10.1371/journal.pone.0093550

Figure Lengend Snippet: Western blot and quantitative densitometry analysis of TLR4-EGFP and β-actin protein expression in each cell line ( A ) and genotype group ( B ). Plotted in panel B are mean TLR4-EGFP/β-actin ratios of densitometry for each genotype cell group. Statistical comparisons were performed using One-way ANOVA and Newman-Keuls post-hoc test among the TLR4 genotype groups (n = 3) except the Asp299Gly/Thr399Ile cell group.

Article Snippet: The following antibodies were used: mouse anti-human TLR4-APC antibody, mouse anti-human TLR4 purified antibody, rat IgG2a K isotype control APC, mouse IgG1 isotype control Alexa Fluor647 and mouse IgG2b isotype control Alexa Fluor647 (eBioscience); rat anti-mouse CD16/CD32, anti-NF-κB p65 antibody (BD Biosciences); PerCP anti-human IL-8 and PerCP mouse Ig G2a isotype control (Biolegend); horseradish peroxidase-conjugated goat anti-rabbit IgG (Invitrogen/LifeTechnologies); rabbit polyclonal antibody against human TLR4 (Santa Cruz Biotechnology, Inc); and rabbit monoclonal antibody against human β-actin (Cell Signaling).

Techniques: Western Blot, Expressing

Journal: PLoS ONE

Article Title: The Toll-Like Receptor 4 Polymorphism Asp299Gly but Not Thr399Ile Influences TLR4 Signaling and Function

doi: 10.1371/journal.pone.0093550

Figure Lengend Snippet: GFP fluorescence, differential interference contrast (DIC) and merged images of 293/Hu-MD2-CD14 cell lines with human WT or polymorphic TLR4-EGFP protein and EGFP control were taken at 400X magnification simultaneously. One representative cell line for each genotype group is displayed. Other cell lines in each group had very similar patterns of TLR4-EGFP expression.

Article Snippet: The following antibodies were used: mouse anti-human TLR4-APC antibody, mouse anti-human TLR4 purified antibody, rat IgG2a K isotype control APC, mouse IgG1 isotype control Alexa Fluor647 and mouse IgG2b isotype control Alexa Fluor647 (eBioscience); rat anti-mouse CD16/CD32, anti-NF-κB p65 antibody (BD Biosciences); PerCP anti-human IL-8 and PerCP mouse Ig G2a isotype control (Biolegend); horseradish peroxidase-conjugated goat anti-rabbit IgG (Invitrogen/LifeTechnologies); rabbit polyclonal antibody against human TLR4 (Santa Cruz Biotechnology, Inc); and rabbit monoclonal antibody against human β-actin (Cell Signaling).

Techniques: Fluorescence, Expressing

Journal: PLoS ONE

Article Title: The Toll-Like Receptor 4 Polymorphism Asp299Gly but Not Thr399Ile Influences TLR4 Signaling and Function

doi: 10.1371/journal.pone.0093550

Figure Lengend Snippet: Shown in panels ( A – D ) are comparisons of dose-response curves to LPS stimulations of cell lines in each TLR4 genotype group. 3.0×10 5 cells were stimulated for 24 hours with a series of LPS concentrations and conditioned media from each cell line was tested for human IL-8 secretion by ELISA. (I) Human IL-8 production 24 hours after LPS treatment for different genotype cell groups. WT-LPS-0 and WT-LPS-20 represent TLR4 wild type cell lines stimulated with 0 and 20 ng/ml LPS (same annotation for other cell groups). ( E – H ) Percentage of TLR4- and EGFP-positive cells that are also positive for IL-8 as assessed by flow cytometry analysis. ( J ) Fold change (percentage multiplied by MFI) of intracellular human IL-8 positive cells after LPS(20 ng/ml)stimulation for the four genotype groups. # no LPS stimulation vs 20 ng/ml LPS stimulation within the same genotypic cell group, P <0.05; ** and *** vs Asp299Gly group, p <0.01 (I) and 0.001(J). Statistical comparisons were performed using One-way ANOVA and Newman-Keuls post-hoc test among the TLR4 genotype groups (n = 3) except the Asp299Gly/Thr399Ile cell group (n = 2).

Article Snippet: The following antibodies were used: mouse anti-human TLR4-APC antibody, mouse anti-human TLR4 purified antibody, rat IgG2a K isotype control APC, mouse IgG1 isotype control Alexa Fluor647 and mouse IgG2b isotype control Alexa Fluor647 (eBioscience); rat anti-mouse CD16/CD32, anti-NF-κB p65 antibody (BD Biosciences); PerCP anti-human IL-8 and PerCP mouse Ig G2a isotype control (Biolegend); horseradish peroxidase-conjugated goat anti-rabbit IgG (Invitrogen/LifeTechnologies); rabbit polyclonal antibody against human TLR4 (Santa Cruz Biotechnology, Inc); and rabbit monoclonal antibody against human β-actin (Cell Signaling).

Techniques: Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: PLoS ONE

Article Title: The Toll-Like Receptor 4 Polymorphism Asp299Gly but Not Thr399Ile Influences TLR4 Signaling and Function

doi: 10.1371/journal.pone.0093550

Figure Lengend Snippet: ( A ) Overlay of percentages of NF-κB phospho-p65 positive cells of one representative cell line in each genotype group after 30 min LPS (10ng/ml) stimulation. Fold changes (percentage of cells multiplied with MFI) of intracellular phosphorylated ( B ) and total ( C ) p65 NF-κB positive cells of different genotype cell groups following 0 min, 30 min, 60 min, and 120 min LPS(10ng/ml)stimulation. Statistical comparisons were performed using One-way ANOVA and Newman-Keuls post-hoc test among the TLR4 genotype groups (n = 3) except the Asp299Gly/Thr399Ile cell group (n = 2).

Article Snippet: The following antibodies were used: mouse anti-human TLR4-APC antibody, mouse anti-human TLR4 purified antibody, rat IgG2a K isotype control APC, mouse IgG1 isotype control Alexa Fluor647 and mouse IgG2b isotype control Alexa Fluor647 (eBioscience); rat anti-mouse CD16/CD32, anti-NF-κB p65 antibody (BD Biosciences); PerCP anti-human IL-8 and PerCP mouse Ig G2a isotype control (Biolegend); horseradish peroxidase-conjugated goat anti-rabbit IgG (Invitrogen/LifeTechnologies); rabbit polyclonal antibody against human TLR4 (Santa Cruz Biotechnology, Inc); and rabbit monoclonal antibody against human β-actin (Cell Signaling).

Techniques: