mouse anti tlr3 monoclonal antibody  (Santa Cruz Biotechnology)

Journal: Animals : an Open Access Journal from MDPI

Article Title: Modulation of the Toll-like Receptor Pathway in Ovine Endometria During Early Pregnancy

doi: 10.3390/ani15070917

Figure Lengend Snippet: Antibodies used for Western blot.

Article Snippet: Mouse anti-TLR3 monoclonal antibody , sc-32232 , Santa Cruz Biotechnology, Santa Cruz, CA, USA , 0.2 μg/mL.

Techniques: Western Blot

Journal: Animals : an Open Access Journal from MDPI

Article Title: Modulation of the Toll-like Receptor Pathway in Ovine Endometria During Early Pregnancy

doi: 10.3390/ani15070917

Figure Lengend Snippet: Relative expression values of ( A ) TLR2 , ( B ) TLR3 , ( C ) TLR4 , ( D ) TLR5 , ( E ) IRAK1 , ( F ) TRAF6 , and ( G ) MYD88 mRNA in the endometrium. Significant differences ( p < 0.05) are indicated by different letters (a, b, c, and d).

Article Snippet: Mouse anti-TLR3 monoclonal antibody , sc-32232 , Santa Cruz Biotechnology, Santa Cruz, CA, USA , 0.2 μg/mL.

Techniques: Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: Modulation of the Toll-like Receptor Pathway in Ovine Endometria During Early Pregnancy

doi: 10.3390/ani15070917

Figure Lengend Snippet: Expression of TLR2, TLR3, TLR4, TLR5, IRAK1, TRAF6, and MyD88 proteins in the endometrium. Significant differences ( p < 0.05) are indicated by different letters (a, b, c, and d).

Article Snippet: Mouse anti-TLR3 monoclonal antibody , sc-32232 , Santa Cruz Biotechnology, Santa Cruz, CA, USA , 0.2 μg/mL.

Techniques: Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: TLR3 signaling in astrocytes. Upon dsRNA recognition in the endosomal compartment, TLR3 undergoes dimerization and interacts with the TRIF adaptor molecule. TRIF activation is followed by TRAF6 and TRAF3 recruitment. TRAF6 conducts the signal via RIP-1 and RIP-3 kinases which facilitate NEMO, IKK- α , and IKK- β complex formation, followed by NF- κ B phosphorylation and translocation into the nucleus. TRAF3 engages TBK1 and IKK-i/ Ɛ for IRF3 and IRF7 activation, followed by their dimerization and translocation into the nucleus. This leads to the induction of type I IFNs and proinflammatory cytokine gene expression. The dotted arrows highlight possible roles of ESCRT-0 in TLR3 transport from the ER to the endosome, as well as the role of Hrs and Syk in TLR3 degradation.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Activation Assay, Translocation Assay, Expressing

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: Primary antibodies used in the western blot assay.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Western Blot, Concentration Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: Poly(I:C) treatment of murine astrocytes induces Syk and Hrs phosphorylation and Syk-Hrs interaction. Hrs interacts with the N-terminal cleaved form of TLR3. (a) After poly(I:C) or poly(I:C)/LyoVec stimulation for 1, 2, 5, 10, and 15 min, the phosphorylation of Syk was analyzed by Western blot. The density level of phosphorylated Syk was normalized to GAPDH. Data was obtained from three independent experiments and presented as mean ± SD. ∗ p ≤ 0.05 and ∗∗ p ≤ 0.01. (b) After poly(I:C) or poly(I:C)/LyoVec stimulation for 5, 8, 12, and 15 min, C8-D1A cells were lysed and Hrs was immunoprecipitated using the anti-Hrs antibody. Phosphotyrosine (P-Tyr), Syk, and TLR3 were then detected by Western blot. (c) Following transfection with Syk siRNA, cells were stimulated with poly(I:C) for 5, 8, 12, and 15 min and lysed and Hrs was immunoprecipitated using the anti-Hrs antibody. Phosphotyrosine (P-Tyr) was detected by Western blot. For all immunoprecipitation experiments, 0 min presents untreated cells and mouse IgG were used as a negative control. EL: immunoprecipitation eluate; FT: immunoprecipitation flow-through; CN: control cell lysate. GAPDH was used as protein loading control. (d) Syk silencing efficiency was visualized by immunoblotting with anti-Syk antibodies.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Western Blot, Immunoprecipitation, Transfection, Negative Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: NF- κ B nuclear translocation is downregulated in poly(I:C)-treated astrocytes with silenced Syk and Hrs. C8-D1A cells were untreated or treated with poly(I:C) or poly(I:C)/LyoVec for 5 min, 8 min, 12 min, 15 min, 30 min, and 60 min. In advance to the stimulation, astrocytes were not transfected (a) or transfected with siRNA pools for TLR3 (b), Syk (c), and Hrs (d). Following poly(I:C) or poly(I:C)/LyoVec treatment, cytoplasmic and nuclear extracts were immunoblotted with anti-NF- κ B p65, -IRF3, -IRF7, -GAPDH, and -PARP antibodies. (e) TLR3 and Hrs silencing efficiency was visualized by immunoblotting with anti-TLR3 and -Hrs antibodies.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Translocation Assay, Transfection, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: Knockdown of Syk and Hrs expression by siRNA upregulates poly(I:C)-induced IFN- β , IL-6, and CXCL-8 production. C8-D1A cells were transfected with control siRNA-A or siRNA pools for TLR3, Syk, Hrs, and STAM. Following the transfection, astrocytes were treated with poly(I:C) (10 μ g/ml) for 24 h. IFN- β (a), IL-6 (b), and CXCL-8 (c) were measured in culture supernatants by ELISA. Because Syk transfection lasted 48 h, in each experiment, supernatants from untreated (not treated (Syk)), poly(I:C)-treated (poly(I:C) (Syk)), and poly(I:C)-treated cells with silenced Syk (siRNA Syk) were tested in the group independent from cells with silenced TLR3, Hrs, and STAM, where transfection lasted 72 h. (d) STAM silencing efficiency was visualized by immunoblotting with anti-STAM antibodies. Data was obtained from three (IFN- β , CXCL-8) or five (IL-6) independent experiments and presented as mean ± SD. ∗ p ≤ 0.05 and ∗∗ p ≤ 0.01.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Expressing, Transfection, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: TLR3 of murine astrocytes is cleaved upon stimulation of cells with poly(I:C). Representative western blots of TLR3 expression in C8D1A cells treated with various concentrations of poly(I:C) (0, 0.1, 1, 2, 5, and 10 μ g/ml) (a) or poly(I:C)-LyoVec (0, 0.1, 1, 2, and 5 μ g/ml) (b) and lysed 24 h after stimulation. TLR3 expression was also analyzed in cells treated with poly(I:C) at concentration 10 μ g/ml (c), or with poly(I:C)-LyoVecat concentration 1 μ g/ml (d), and lysed at various times of stimulation (0, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h). TLR3 FL: full-length TLR3; TLR3 N: cleaved N-terminal TLR3 form; GAPDH: protein loading control. Densitometry analysis of TLR3 forms was performed in cells treated with indicated poly(I:C) concentrations for 24 h (a), indicated poly(I:C)/LyoVecconcentrations for 24 h (b), 10 μ g/ml poly(I:C) for indicated time points (c), or 1 μ g/ml poly(I:C)/LyoVec for indicated time points (d). The density level of each protein was normalized to GAPDH. Data was obtained from three independent experiments and presented as mean ± SD. ∗ p ≤ 0.05 and ∗∗ p ≤ 0.01.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Western Blot, Expressing, Concentration Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: Stimulation of murine astrocytes with poly(I:C) leads to the time-dependent increase in Syk and Hrs expression, while the expression of STAM does not significantly change after stimulation of cells with the TLR3 ligand. Representative western blots of Hrs, Syk, and STAM expression in C8D1A cells treated with various concentrations of poly(I:C) (0, 0.1, 1, 2, 5, and 10 μ g/ml) (a), or poly(I:C)-LyoVec (0, 0.1, 1, 2, and 5 μ g/ml) (b), and lysed 24 h after stimulation. Hrs, Syk, and STAM expression was also analyzed in cells treated with poly(I:C) at concentration 10 μ g/ml (c), or with poly(I:C)-LyoVecat concentration 1 μ g/ml (d), and lysed at various times of stimulation (0, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h). GAPDH was used for evaluating protein loading control. Densitometry analysis of Hrs, Syk, and STAM was performed in cells treated with indicated poly(I:C) concentrations for 24 h (a), indicated poly(I:C)/LyoVecconcentrations for 24 h (b), 10 μ g/ml poly(I:C) for indicated time points (c), or 1 μ g/ml poly(I:C)/LyoVec for indicated time points (d). The density level of each protein was normalized to GAPDH. Data was obtained from three independent experiments and presented as mean ± SD. ∗ p ≤ 0.05 and ∗∗ p ≤ 0.01.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Expressing, Western Blot, Concentration Assay

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: Immunostains of TLR3, Syk, Hrs, and STAM expression and localization in C8D1A cells after treatment with poly(I:C). C8-D1A murine astrocytes were not treated or treated with poly(I:C) (10 μ g/ml) for 5 min, 4 h, and 24 h, fixed and immunostained with specific antibodies. Selected images present intracellular distribution of TLR3 (a), Syk (b), Hrs (c), and STAM (d) (red fluorescence). To visualize colocalization of ER with TLR3 or STAM, following poly(I:C) stimulation at the indicated time points, cells were double stained with anti-TLR3 (red) and anti-PDI (green) antibodies (e), or with anti-STAM (red) and anti-PDI (green) antibodies (f). Nuclear DNA was stained with Hoechst 33342 (blue fluorescence). Scale bar = 10 μ m, N.T. = no treatment.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Expressing, Fluorescence, Staining

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Syk and Hrs Regulate TLR3-Mediated Antiviral Response in Murine Astrocytes

doi: 10.1155/2019/6927380

Figure Lengend Snippet: Poly(I:C) treatment of murine astrocytes induces TLR3 tyrosine phosphorylation and promotes interaction with Hrs. (a) After poly(I:C) or poly(I:C)/LyoVec stimulation for 5, 8, 12, 15, and 30 min, C8-D1A cells were lysed and TLR3 was immunoprecipitated using the anti-TLR3 antibody. Phosphotyrosine (P-Tyr), Hrs, and STAM were then detected by Western blot. (b) Following poly(I:C) or poly(I:C)/LyoVec stimulation for 5, 8, 12, andn 15 min, C8-D1A cells were lysed and TLR3 was immunoprecipitated using the anti-TLR3 antibody. Ubiquitin was detected by Western blot. Blue arrows indicate ubiquitinated TLR3. (c) Following poly(I:C) stimulation for 5, 8, 12, and 15 min, murine astrocytes were lysed and STAM and Hrs were immunoprecipitated using anti-STAM and anti-Hrs antibodies, respectively. Hrs and STAM were detected by Western blot. For all immunoprecipitation experiments, 0 min presents untreated cells and mouse IgG were used as a negative control. EL: immunoprecipitation eluate; FT: immunoprecipitation flow through; CN: control cell lysate. GAPDH was used as protein loading control.

Article Snippet: TLR3 , TLR3.7 b , Monoclonal mouse , OriGene Technologies GmbH , 2 μ g/ml.

Techniques: Immunoprecipitation, Western Blot, Negative Control

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: Oligonucleotide primers used for the quantitative real time PCR.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques:

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 transcript are expressed in endometrium during the menstrual cycle . Columns indicate mean TLR3 and TLR4 mRNA quantities from endometrium in proliferative (n = 16), secretory (n = 11) and menstrual phase (n = 8) run in triplicates. The y-axis is scaled logarithmically; error bars represent the standard deviation of the mean.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Standard Deviation

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 protein is localised to endometrial cells during the menstrual cycle . TLR3 protein staining in healthy late proliferative (LP) tissue was high in luminal and glandular tissue (A , brown precipitate) and lower in LP endometriotic tissue (B) . Late secretory (LS) endometrium showed highly expressed TLR3 in the epithelium (C) , but weakly in endometriosis (D) . Intense staining of TLR4 proteins was shown in mid proliferative (MP) tissue (E) . In late proliferative phase of endometriosis, TLR4 proteins were comparably lower (F) . TLR4 protein was high in mid secretory (MS) normal endometrium (G) , whereas it was decreased in endometriotic MS tissue (H) . During the menstrual phase, both TLR3 (I) and TLR4 (J) were highly expressed. Co-immunostaining for TLR4 (green), CD14 ( K , red) and CD163 ( L , red) demonstrated that TLR4 proteins were expressed by CD14 positive dendritic cells and monocytes ( K , yellow) and by CD163 positive macrophages ( L , yellow). Localisation of TLR4 to immune cells is marked by a black arrow (J) and by white arrows (K, L) .

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Staining, Immunostaining

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 mRNA expression is regulated in endometriosis . The expression of TLR3 (A, C) and TLR4 mRNA (B, D) in endometrium during proliferative (n = 13, run in triplicates, A, B) and secretory phase (n = 3, C, D) was decreased in eutopic endometriotic endometrium when compared to controls. In addition, four proliferative corresponding lesions were evaluated (A, B) showing a local upregulation of both receptors on ectopic sites. Columns represent the mean ratio of TLR copy number to ACTB copy number. Error bars represent the standard deviation of the mean. * P < 0.05; ** P < 0.01.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Expressing, Standard Deviation

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 protein is locally induced in endometriotic lesions . No specific TLR3 protein staining was seen in eutopic endometriotic tissue (A) whereas a high glandular localisation of the protein was detected in a gland of an ectopic endometriotic lesion from the same patient (B) . Similarly, TLR4 was not detectable in eutopic endometrium (C) but present in glandular epithelium of ectopic endometrium from the same women (D) .

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Staining

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 mRNA expression is decreased in endometrial adenocarcinoma . (A-B) Columns indicate mean TLR3 (A) and TLR4 (B) mRNA levels from postmenopausal patients (PMP, n = 8), and those diagnosed with endometrial hyperplasia (HP, n = 10) and endometrial carcinoma (EnCa, n = 16). (C-D) TLR3 (C) and TLR4 (D) mRNA expression in different carcinoma grades compared to postmenopausal controls and hyperplastic endometrium: G1 (n = 5), G2 (n = 6) and G3 (n = 5). Error bars represent the standard deviation of the mean. * P < 0.05; ** P < 0.01.

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Expressing, Standard Deviation

Journal: Reproductive biology and endocrinology : RB&E

Article Title: TLR3 and TLR4 expression in healthy and diseased human endometrium

doi: 10.1186/1477-7827-6-40

Figure Lengend Snippet: TLR3 and TLR4 proteins are present in postmenopausal endometrium . Localisation of TLR3 in normal postmenopausal endometrium (A) , endometrial hyperplasia (B) , endometrial adenocarcinoma grade G1 (C) , G2 (D) and G3 (E ). Localisation of TLR4 protein in normal postmenopausal endometrium (F) , endometrial hyperplasia (G) , endometrial adenocarcinoma grade G1 (H) , G2 (I) and G3( J ). All stained sections indicated epithelium as the preferred localisation of TLR3 and TLR4 proteins. TLR4 protein was additionally present in immune cells (arrows).

Article Snippet: Slides were incubated in a humidified chamber overnight at 4°C with the monoclonal mouse-anti-human antibodies against TLR3 [ ] and TLR4 [HTA125, [ ]] at 20 μg/ml and 100 μg/ml, respectively (Acris Antibodies, Hiddenhausen, Germany).

Techniques: Staining

Journal: Immunologic Research

Article Title: Tryptanthrin attenuates TLR3-mediated STAT1 activation in THP-1 cells

doi: 10.1007/s12026-022-09301-z

Figure Lengend Snippet: A Cells were incubated with 1 μM tryptanthrin or DMSO for 24 h, and the total RNA was extracted. The mRNA expression of toll-like receptor 3 (TLR3) was examined by qRT-PCR ( n = 3; n.s., not significant). B Flow cytometry analysis of TLR3 expression in THP-1 Mϕ treated with 1 μM tryptanthrin (right panel) or DMSO (left panel). Representative histograms of three to five independent experiments. C Quantification of mean fluorescence intensity for TLR3 ( n = 3; n.s., not significant). D Cells were incubated with 1 μM tryptanthrin or DMSO for 24 h, fixed with methanol, and immunostained with anti-TLR3 antibody (green). Cell nuclei were stained using DAPI (blue). Representative images of three independent experiments. E , F Cells were stimulated with 1 ng/mL rIFN-β ( E ) or rIFN-γ ( F ) with or without tryptanthrin for up to 30 min, and the cell lysates were subjected to western blotting to analyze p-STAT1-Tyr and STAT1. G Putative mechanism of tryptanthrin-mediated regulation of TLR3 signaling in THP Mϕ. Tryptanthrin negatively modulates IRF3 activation and the subsequent IFN-β-induced phosphorylation of STAT1

Article Snippet: Mouse monoclonal anti-IRF3 (sc-33641), rabbit polyclonal anti-STAT1 (sc-346), mouse monoclonal anti-p-STAT1 (pY701.4A) (p-STAT1-Tyr) (sc-136229), and mouse monoclonal anti-TLR3 (sc-32232) antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Incubation, Expressing, Quantitative RT-PCR, Flow Cytometry, Fluorescence, Staining, Western Blot, Activation Assay, Phospho-proteomics