rabbit polyclonal anti-mouse tlr2  (Thermo Fisher)

Journal: Frontiers in Pharmacology

Article Title: Oleanolic Acid Acetate Alleviates Symptoms of Experimental Autoimmune Encephalomyelitis in Mice by Regulating Toll-Like Receptor 2 Signaling

doi: 10.3389/fphar.2020.556391

Figure Lengend Snippet: OAA suppressed infiltration of immune cells and macrophages in the spinal cords of EAE mice. H&E staining showed that OAA administration attenuated inflammatory infiltration in spinal cords compared to the infiltration observed in EAE mice (upper panel). Immunohistochemistry showed that infiltration of CD68+ macrophages was reduced in the spinal cords of OAA-treated mice (lower panel). Expression of TLR2 was elevated in EAE spinal cords and mainly expressed in CD68 + macrophages (lower panel). Scale bars = 100 µm. OAA, oleanolic acid acetate; EAE, experimental autoimmune encephalomyelitis; H&E, hematoxylin and eosin.

Article Snippet: The slides were incubated overnight with rabbit polyclonal anti-mouse TLR2 (1:100, Thermo Fisher Scientific Inc., Waltham, MA, USA), and rat monoclonal anti-mouse CD68 (1:100, Thermo Fisher Scientific Inc., Waltham, MA, USA) antibodies at 4°C.

Techniques: Staining, Immunohistochemistry, Expressing

Journal: Frontiers in Pharmacology

Article Title: Oleanolic Acid Acetate Alleviates Symptoms of Experimental Autoimmune Encephalomyelitis in Mice by Regulating Toll-Like Receptor 2 Signaling

doi: 10.3389/fphar.2020.556391

Figure Lengend Snippet: OAA downregulated the expression of TLR2 and downstream signaling molecules in the spinal cords of EAE mice. OAA downregulated TLR2 and MyD88 mRNA expression in the spinal cords (A) . Western blot analysis of TLR2, MyD88, IRAK4, and TRAF6 (B) . The expression level of TLR2, MyD88, IRAK4, and TRAF6 was significantly suppressed in OAA treated group, compared with that of EAE group (B) . Data are shown as mean ± standard error (SE), n=6, *p < 0.05 compared to EAE mice. OAA, oleanolic acid acetate; EAE, experimental autoimmune encephalomyelitis; TLR2, Toll-like receptor 2; MyD88, myeloid differentiation primary response protein 88; IRAK4, IL-1 receptor-associated kinase 4: TRAF6, tumor necrosis factor (TNF) receptor-associated factor 6.

Article Snippet: The slides were incubated overnight with rabbit polyclonal anti-mouse TLR2 (1:100, Thermo Fisher Scientific Inc., Waltham, MA, USA), and rat monoclonal anti-mouse CD68 (1:100, Thermo Fisher Scientific Inc., Waltham, MA, USA) antibodies at 4°C.

Techniques: Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: (A–F) Expression of TLR2 in lung tissue from the mice in each group was assessed using immunohistochemistry (deep brown cellular area indicates positive staining; magnification, x200). (G) Quantification of TLR2-positive staining in five fields of view per section from each mouse was assessed by JD-801 computer-aided image analyzer under high-power magnification (x200). Values are expressed as the mean ± standard deviation of five mice in each group. * P<0.05, as compared with the NC group. # P<0.05, as compared with the AM group. NC, normal control; AM, ovalbumin immunization-induced asthmatic mice; BAM, BML-111-treated AM; VAM, vehicle of BML-111-treated AM; AAM, anti-IL-1β antibody-treated AM; RAM, rabbit IgG-treated AM; TLR, of toll-like receptor.

Article Snippet: The blots were incubated with primary rabbit anti-mouse polyclonal antibodies against TLR2 (cat. no. sc-10739), MyD88 (cat. no. sc-11356) or β-actin (cat. no. sc-130656; Santa Cruz Biotechnology, Inc.) at 1:2,000 dilution overnight followed by incubation for 2 h with an HRP-conjugated goat anti-rabbit IgG antibody (Jackson, West Grove, PA, USA) at 1:5,000 dilution.

Techniques: Expressing, Immunohistochemistry, Staining, Standard Deviation, Control

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Expressions of TLR2 and MyD88 assessed using western blot analysis in lung tissue obtained from the mice in each group. The western blot shown is a representative of five independent experiments, and β-actin protein served as a loading control. Semiquantitative analysis was performed by using UVP-gel densitometry. Arbitrary unit = (A TLR2 /A β-actin ) × 100%, or (A MyD88 /A β-actin ) ×100%. Values are expressed as mean ± standard deviation of five mice in each group. * P<0.05, as compared with the arbitrary units of the same protein in the NC group. # P<0.05, as compared with the arbitrary units of the same protein in the AM group. NC, normal control; AM, ovalbumin immunization-induced asthmatic mice; BAM, BML-111-treated AM; VAM, vehicle of BML-111-treated AM; AAM, anti-IL-1β antibody-treated AM; RAM, rabbit immunoglobulin G-treated AM; MyD88, mycloid differentiation factor 88; TLR, toll-like receptor.

Article Snippet: The blots were incubated with primary rabbit anti-mouse polyclonal antibodies against TLR2 (cat. no. sc-10739), MyD88 (cat. no. sc-11356) or β-actin (cat. no. sc-130656; Santa Cruz Biotechnology, Inc.) at 1:2,000 dilution overnight followed by incubation for 2 h with an HRP-conjugated goat anti-rabbit IgG antibody (Jackson, West Grove, PA, USA) at 1:5,000 dilution.

Techniques: Western Blot, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Supernatant levels of IL-4, IL-6 and IL-8 released from leukocytes exposed to IL-1β were assessed using ELISA. The cultured leukocytes were obtained from a mouse from the normal control group and stimulated with IL-1β (10 ng/ml) for 24 h with or without pre-treatment of BML-111 (1 mM) for 30 min, antagonist of G protein-coupled LXA4 receptor BOC1 (100 μ M) for 30 min, TLR2Ab (1 μ g/ml) for 1 h, goat IgG (1 μ g/ml) for 1 h, MyD88 dimerization inhibitor ST2825 (20 μ M) for 1 h and TPCA-1 (10 μ M) for 1 h. Values are expressed as the mean ± standard deviation of five independent experiments. * P<0.05, as compared to the levels of same cytokine released from the cells without treatment. # P<0.05, as compared to the levels of same cytokine released from the cells treated with IL-1β alone. IL, interleukin; IgG, immunoglobulin G; TLR, toll-like receptor; TPCA-1,10, thiophene-3-carboxamide 1; TLR2Ab, toll-like receptor 2-neutralizing antibody; BOC1, N -t-Boc-Phe-Leu-Phe-Leu-Phe.

Article Snippet: The blots were incubated with primary rabbit anti-mouse polyclonal antibodies against TLR2 (cat. no. sc-10739), MyD88 (cat. no. sc-11356) or β-actin (cat. no. sc-130656; Santa Cruz Biotechnology, Inc.) at 1:2,000 dilution overnight followed by incubation for 2 h with an HRP-conjugated goat anti-rabbit IgG antibody (Jackson, West Grove, PA, USA) at 1:5,000 dilution.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Expression of TLR2 and MyD88 assessed using western blot analysis of leukocytes exposed to IL-1β. The cultured leukocytes were obtained from a normal control mouse and stimulated with IL-1β (10 ng/ml) for 30 min with or without pre-treatment of BML-111 (1 mM) for 30 min, antagonist of G protein-coupled LXA4 receptor BOC1 (100 μ M) for 30 min, TLR2Ab (1 μ g/ml) for 1 h, goat IgG (1 μ g/ml) for 1 h and MyD88 dimerization inhibitor ST2825 (20 μ M) for 1 h. The western blot is representative of five independent experiments, and the lower panel shows β-actin protein, which served as a loading control. Semiquantitative analysis was performed by using UVP-gel densitometry. Arbitrary unit = (A TLR2 /A β-actin ) ×100%, or (A MyD88 /A β-actin ) ×100%. Values are expressed as the mean ± standard deviation of five independent experiments. * P<0.05, as compared to the arbitrary units of the same protein in the cells without treatment. # P<0.05, as compared to the arbitrary units of the same protein in the cells treated with IL-1β alone. IL, interleukin; IgG, immunoglobulin; TLR, toll-like receptor; TLR2Ab, toll-like receptor 2-neutralizing antibody; BOC1, N -t-Boc-Phe-Leu-Phe-Leu-Phe.

Article Snippet: The blots were incubated with primary rabbit anti-mouse polyclonal antibodies against TLR2 (cat. no. sc-10739), MyD88 (cat. no. sc-11356) or β-actin (cat. no. sc-130656; Santa Cruz Biotechnology, Inc.) at 1:2,000 dilution overnight followed by incubation for 2 h with an HRP-conjugated goat anti-rabbit IgG antibody (Jackson, West Grove, PA, USA) at 1:5,000 dilution.

Techniques: Expressing, Western Blot, Cell Culture, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Activation of NF-κB assessed using western blot analysis in leukocytes exposed to IL-1β. The cultured leukocytes were obtained from a normal control mouse and stimulated with IL-1β (10 ng/ml) for 1 h with or without pre-treatment of BML-111 (1 mM) for 30 min, BOC1 (100 μ M) for 30 min, TLR2Ab (1 μ g/ml) for 1 h, goat IgG (1 μ g/ml) for 1 h, ST2825 (20 μ M) for 1 h or TPCA-1 (10 μ M) for 1 h. The western blot is representative of five independent experiments. GAPDH served as a loading control of cytosolic proteins. Histone 3 protein served as a loading control of nuclear proteins. Semiquantitative analysis was performed using UVP-gel densitometry. Arbitrary unit = (A IκBα /A GAPDH ) ×100%, or (A cytosolic p65 /A GAPDH ) ×100%, or (A nuclear p65 /A histone 3 ) ×100%. Values are expressed as the mean ± standard deviation of five independent experiments. * P<0.05, as compared to the arbitrary units of the same protein in the cells without treatment. IL, interleukin; IgG, immunoglobulin G; TPCA-1,10, thiophene-3-carbox-amide 1; TLR2Ab, toll-like receptor 2-neutralizing antibody; BOC1, N -t-Boc-Phe-Leu-Phe-Leu-Phe; NF-κB, nuclear factor-κB; IκBα, inhibitor of kappa B alpha.

Article Snippet: The blots were incubated with primary rabbit anti-mouse polyclonal antibodies against TLR2 (cat. no. sc-10739), MyD88 (cat. no. sc-11356) or β-actin (cat. no. sc-130656; Santa Cruz Biotechnology, Inc.) at 1:2,000 dilution overnight followed by incubation for 2 h with an HRP-conjugated goat anti-rabbit IgG antibody (Jackson, West Grove, PA, USA) at 1:5,000 dilution.

Techniques: Activation Assay, Western Blot, Cell Culture, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: (A–F) Expression of TLR2 in lung tissue from the mice in each group was assessed using immunohistochemistry (deep brown cellular area indicates positive staining; magnification, x200). (G) Quantification of TLR2-positive staining in five fields of view per section from each mouse was assessed by JD-801 computer-aided image analyzer under high-power magnification (x200). Values are expressed as the mean ± standard deviation of five mice in each group. * P<0.05, as compared with the NC group. # P<0.05, as compared with the AM group. NC, normal control; AM, ovalbumin immunization-induced asthmatic mice; BAM, BML-111-treated AM; VAM, vehicle of BML-111-treated AM; AAM, anti-IL-1β antibody-treated AM; RAM, rabbit IgG-treated AM; TLR, of toll-like receptor.

Article Snippet: Sections were incubated with either rabbit anti-mouse polyclonal TLR2 antibody (cat. no. sc-10739) or rabbit anti-mouse polyclonal phosphorylated NF-κB p65 (Ser 276) antibody (cat. no. sc-109; Santa Cruz Biotechnology, Inc.) at 1:100 dilution.

Techniques: Expressing, Immunohistochemistry, Staining, Standard Deviation, Control

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Expressions of TLR2 and MyD88 assessed using western blot analysis in lung tissue obtained from the mice in each group. The western blot shown is a representative of five independent experiments, and β-actin protein served as a loading control. Semiquantitative analysis was performed by using UVP-gel densitometry. Arbitrary unit = (A TLR2 /A β-actin ) × 100%, or (A MyD88 /A β-actin ) ×100%. Values are expressed as mean ± standard deviation of five mice in each group. * P<0.05, as compared with the arbitrary units of the same protein in the NC group. # P<0.05, as compared with the arbitrary units of the same protein in the AM group. NC, normal control; AM, ovalbumin immunization-induced asthmatic mice; BAM, BML-111-treated AM; VAM, vehicle of BML-111-treated AM; AAM, anti-IL-1β antibody-treated AM; RAM, rabbit immunoglobulin G-treated AM; MyD88, mycloid differentiation factor 88; TLR, toll-like receptor.

Article Snippet: Sections were incubated with either rabbit anti-mouse polyclonal TLR2 antibody (cat. no. sc-10739) or rabbit anti-mouse polyclonal phosphorylated NF-κB p65 (Ser 276) antibody (cat. no. sc-109; Santa Cruz Biotechnology, Inc.) at 1:100 dilution.

Techniques: Western Blot, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Supernatant levels of IL-4, IL-6 and IL-8 released from leukocytes exposed to IL-1β were assessed using ELISA. The cultured leukocytes were obtained from a mouse from the normal control group and stimulated with IL-1β (10 ng/ml) for 24 h with or without pre-treatment of BML-111 (1 mM) for 30 min, antagonist of G protein-coupled LXA4 receptor BOC1 (100 μ M) for 30 min, TLR2Ab (1 μ g/ml) for 1 h, goat IgG (1 μ g/ml) for 1 h, MyD88 dimerization inhibitor ST2825 (20 μ M) for 1 h and TPCA-1 (10 μ M) for 1 h. Values are expressed as the mean ± standard deviation of five independent experiments. * P<0.05, as compared to the levels of same cytokine released from the cells without treatment. # P<0.05, as compared to the levels of same cytokine released from the cells treated with IL-1β alone. IL, interleukin; IgG, immunoglobulin G; TLR, toll-like receptor; TPCA-1,10, thiophene-3-carboxamide 1; TLR2Ab, toll-like receptor 2-neutralizing antibody; BOC1, N -t-Boc-Phe-Leu-Phe-Leu-Phe.

Article Snippet: Sections were incubated with either rabbit anti-mouse polyclonal TLR2 antibody (cat. no. sc-10739) or rabbit anti-mouse polyclonal phosphorylated NF-κB p65 (Ser 276) antibody (cat. no. sc-109; Santa Cruz Biotechnology, Inc.) at 1:100 dilution.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Expression of TLR2 and MyD88 assessed using western blot analysis of leukocytes exposed to IL-1β. The cultured leukocytes were obtained from a normal control mouse and stimulated with IL-1β (10 ng/ml) for 30 min with or without pre-treatment of BML-111 (1 mM) for 30 min, antagonist of G protein-coupled LXA4 receptor BOC1 (100 μ M) for 30 min, TLR2Ab (1 μ g/ml) for 1 h, goat IgG (1 μ g/ml) for 1 h and MyD88 dimerization inhibitor ST2825 (20 μ M) for 1 h. The western blot is representative of five independent experiments, and the lower panel shows β-actin protein, which served as a loading control. Semiquantitative analysis was performed by using UVP-gel densitometry. Arbitrary unit = (A TLR2 /A β-actin ) ×100%, or (A MyD88 /A β-actin ) ×100%. Values are expressed as the mean ± standard deviation of five independent experiments. * P<0.05, as compared to the arbitrary units of the same protein in the cells without treatment. # P<0.05, as compared to the arbitrary units of the same protein in the cells treated with IL-1β alone. IL, interleukin; IgG, immunoglobulin; TLR, toll-like receptor; TLR2Ab, toll-like receptor 2-neutralizing antibody; BOC1, N -t-Boc-Phe-Leu-Phe-Leu-Phe.

Article Snippet: Sections were incubated with either rabbit anti-mouse polyclonal TLR2 antibody (cat. no. sc-10739) or rabbit anti-mouse polyclonal phosphorylated NF-κB p65 (Ser 276) antibody (cat. no. sc-109; Santa Cruz Biotechnology, Inc.) at 1:100 dilution.

Techniques: Expressing, Western Blot, Cell Culture, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Roles of lipoxin A4 receptor activation and anti-interleukin-1β antibody on the toll-like receptor 2/mycloid differentiation factor 88/nuclear factor-κB pathway in airway inflammation induced by ovalbumin

doi: 10.3892/mmr.2015.3443

Figure Lengend Snippet: Activation of NF-κB assessed using western blot analysis in leukocytes exposed to IL-1β. The cultured leukocytes were obtained from a normal control mouse and stimulated with IL-1β (10 ng/ml) for 1 h with or without pre-treatment of BML-111 (1 mM) for 30 min, BOC1 (100 μ M) for 30 min, TLR2Ab (1 μ g/ml) for 1 h, goat IgG (1 μ g/ml) for 1 h, ST2825 (20 μ M) for 1 h or TPCA-1 (10 μ M) for 1 h. The western blot is representative of five independent experiments. GAPDH served as a loading control of cytosolic proteins. Histone 3 protein served as a loading control of nuclear proteins. Semiquantitative analysis was performed using UVP-gel densitometry. Arbitrary unit = (A IκBα /A GAPDH ) ×100%, or (A cytosolic p65 /A GAPDH ) ×100%, or (A nuclear p65 /A histone 3 ) ×100%. Values are expressed as the mean ± standard deviation of five independent experiments. * P<0.05, as compared to the arbitrary units of the same protein in the cells without treatment. IL, interleukin; IgG, immunoglobulin G; TPCA-1,10, thiophene-3-carbox-amide 1; TLR2Ab, toll-like receptor 2-neutralizing antibody; BOC1, N -t-Boc-Phe-Leu-Phe-Leu-Phe; NF-κB, nuclear factor-κB; IκBα, inhibitor of kappa B alpha.

Article Snippet: Sections were incubated with either rabbit anti-mouse polyclonal TLR2 antibody (cat. no. sc-10739) or rabbit anti-mouse polyclonal phosphorylated NF-κB p65 (Ser 276) antibody (cat. no. sc-109; Santa Cruz Biotechnology, Inc.) at 1:100 dilution.

Techniques: Activation Assay, Western Blot, Cell Culture, Control, Standard Deviation