rabbit anti mouse tlr4 polyclonal antibody  (Novus Biologicals)

Journal: Pharmaceutical biology

Article Title: Berberine attenuates septic cardiomyopathy by inhibiting TLR4/NF-κB signalling in rats.

doi: 10.1080/13880209.2021.1877736

Figure Lengend Snippet: Figure 3. Effects of berberine on key effectors of the TLR4/NF-jB signalling pathway. (A) TLR4 protein expression in the myocardial tissue and quantitation. (B) Nuclear p65 protein expression in the rat sepsis cardiomyopathy model measured by ELISA. Effects of berberine on TNFa (C) and IL 1b (D) levels in the myocardial tis- sue. p < 0.001 indicates significance in comparison with the Con group. p < 0.001 indicates significance in comparison with the LPS group.

Article Snippet: The membranes were blocked with 5% skim milk, followed by successive incubations with rabbit anti-mouse TLR4 polyclonal antibody (1:2000, Santa Cruz, USA) at 4 C overnight and mouse anti-rabbit IgG monoclonal secondary antibody (1:10000, Bioss, China) for 2h.

Techniques: Expressing, Quantitation Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-16 regulates lipopolysaccharide-induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

doi: 10.3892/etm.2021.10414

Figure Lengend Snippet: miR-16 directly targets TLR4. (A) A specific binding site of miR-16 was identified within the 3'-UTR of TLR4 using Targetscan bioinformatics analysis. (B) Luciferase reporter assay was used to verify if miR-16 could directly target the TLR4 3'-UTR in 293T cells. (C) Western blot analysis for TLR4 protein expression after miR-16 mimic transfection in NHBE cells. * P<0.05 (compare with wild NC group). 3'-UTR, 3'-untranslated region; TLR4, toll-like receptor 4; miR, microRNA; NC, negative control.

Article Snippet: The membranes were first probed with the anti-TLR4 mouse polyclonal antibody incubated at 4˚C overnight (cat. no. sc-293072; dilution 1:1,500; Santa Cruz Biotechnology, Inc.) and then with a corresponding horseradish peroxidase-conjugated secondary antibody (cat. no. sc-525409; 1:5,000; Santa Cruz Biotechnology, Inc.) at room temperature for 1.5 h. An enhanced chemiluminescence kit (EMD Millipore) was used for visualization of the results and quantification of the bands was performed using the Image J software (v1.8.0; National Institutes of Health).

Techniques: Binding Assay, Luciferase, Reporter Assay, Western Blot, Expressing, Transfection, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-16 regulates lipopolysaccharide-induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

doi: 10.3892/etm.2021.10414

Figure Lengend Snippet: TLR4 knockdown alleviates LPS-induced inflammatory factors secretion from NHBE cells. (A) Western blot analysis for TLR4 protein expression after transfection with si-TLR4. The secretion levels of (B) TNF-α, (C) IL-1β and (D) IL-6 were measured by ELISA following transfection of NHBE cells with si-TLR4 or si-NC for 48 h. * P<0.05. LPS, lipopolysaccharide; TNF-α, tumor necrosis factor α; IL, interleukin; si, small interfering; NC, negative control; TLR4, toll-like receptor 4; NHBE, normal human bronchial epithelial.

Article Snippet: The membranes were first probed with the anti-TLR4 mouse polyclonal antibody incubated at 4˚C overnight (cat. no. sc-293072; dilution 1:1,500; Santa Cruz Biotechnology, Inc.) and then with a corresponding horseradish peroxidase-conjugated secondary antibody (cat. no. sc-525409; 1:5,000; Santa Cruz Biotechnology, Inc.) at room temperature for 1.5 h. An enhanced chemiluminescence kit (EMD Millipore) was used for visualization of the results and quantification of the bands was performed using the Image J software (v1.8.0; National Institutes of Health).

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

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-16 regulates lipopolysaccharide-induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

doi: 10.3892/etm.2021.10414

Figure Lengend Snippet: miR-16 exerts protective effects against ALI-induced inflammatory responses by modulating TLR4 expression. Secretion levels of (A) TNF-α, (B) IL-1β and (C) IL-6 were assessed by ELISA after co-transfection and LPS treatment. * P<0.05. ALI, acute lung injury; TNF-α, tumor necrosis factor α; IL-1β, interleukin 1β.

Article Snippet: The membranes were first probed with the anti-TLR4 mouse polyclonal antibody incubated at 4˚C overnight (cat. no. sc-293072; dilution 1:1,500; Santa Cruz Biotechnology, Inc.) and then with a corresponding horseradish peroxidase-conjugated secondary antibody (cat. no. sc-525409; 1:5,000; Santa Cruz Biotechnology, Inc.) at room temperature for 1.5 h. An enhanced chemiluminescence kit (EMD Millipore) was used for visualization of the results and quantification of the bands was performed using the Image J software (v1.8.0; National Institutes of Health).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cotransfection

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-16 regulates lipopolysaccharide-induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

doi: 10.3892/etm.2021.10414

Figure Lengend Snippet: miR-16 directly targets TLR4. (A) A specific binding site of miR-16 was identified within the 3'-UTR of TLR4 using Targetscan bioinformatics analysis. (B) Luciferase reporter assay was used to verify if miR-16 could directly target the TLR4 3'-UTR in 293T cells. (C) Western blot analysis for TLR4 protein expression after miR-16 mimic transfection in NHBE cells. * P<0.05 (compare with wild NC group). 3'-UTR, 3'-untranslated region; TLR4, toll-like receptor 4; miR, microRNA; NC, negative control.

Article Snippet: The membranes were first probed with the anti-TLR4 mouse polyclonal antibody incubated at 4 ̊C overnight (cat. no. sc-293072; dilution 1:1,500; Santa Cruz Biotechnology, Inc.) and then with a corresponding horseradish peroxidase-conjugated secondary antibody (cat. no. sc-525409; 1:5,000; Santa Cruz Biotechnology, Inc.) at room temperature for 1.5 h. An enhanced chemiluminescence kit (EMD Millipore) was used for visualization of the results and quantification of the bands was performed using the Image J software (v1.8.0; National Institutes of Health).

Techniques: Binding Assay, Luciferase, Reporter Assay, Western Blot, Expressing, Transfection, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-16 regulates lipopolysaccharide-induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

doi: 10.3892/etm.2021.10414

Figure Lengend Snippet: TLR4 knockdown alleviates LPS-induced inflammatory factors secretion from NHBE cells. (A) Western blot analysis for TLR4 protein expression after transfection with si-TLR4. The secretion levels of (B) TNF-α, (C) IL-1β and (D) IL-6 were measured by ELISA following transfection of NHBE cells with si-TLR4 or si-NC for 48 h. * P<0.05. LPS, lipopolysaccharide; TNF-α, tumor necrosis factor α; IL, interleukin; si, small interfering; NC, negative control; TLR4, toll-like receptor 4; NHBE, normal human bronchial epithelial.

Article Snippet: The membranes were first probed with the anti-TLR4 mouse polyclonal antibody incubated at 4 ̊C overnight (cat. no. sc-293072; dilution 1:1,500; Santa Cruz Biotechnology, Inc.) and then with a corresponding horseradish peroxidase-conjugated secondary antibody (cat. no. sc-525409; 1:5,000; Santa Cruz Biotechnology, Inc.) at room temperature for 1.5 h. An enhanced chemiluminescence kit (EMD Millipore) was used for visualization of the results and quantification of the bands was performed using the Image J software (v1.8.0; National Institutes of Health).

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

Journal: Experimental and Therapeutic Medicine

Article Title: MicroRNA-16 regulates lipopolysaccharide-induced inflammatory factor expression by targeting TLR4 in normal human bronchial epithelial cells

doi: 10.3892/etm.2021.10414

Figure Lengend Snippet: miR-16 exerts protective effects against ALI-induced inflammatory responses by modulating TLR4 expression. Secretion levels of (A) TNF-α, (B) IL-1β and (C) IL-6 were assessed by ELISA after co-transfection and LPS treatment. * P<0.05. ALI, acute lung injury; TNF-α, tumor necrosis factor α; IL-1β, interleukin 1β.

Article Snippet: The membranes were first probed with the anti-TLR4 mouse polyclonal antibody incubated at 4 ̊C overnight (cat. no. sc-293072; dilution 1:1,500; Santa Cruz Biotechnology, Inc.) and then with a corresponding horseradish peroxidase-conjugated secondary antibody (cat. no. sc-525409; 1:5,000; Santa Cruz Biotechnology, Inc.) at room temperature for 1.5 h. An enhanced chemiluminescence kit (EMD Millipore) was used for visualization of the results and quantification of the bands was performed using the Image J software (v1.8.0; National Institutes of Health).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Cotransfection