rabbit polyclonal anti tlr2 antibody  (Biorbyt)

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: Temporal expression of TLR2 in rat brain after DAI. (a) A coronal brain map showed the region where the TLR2 expression was observed in frontal cortex, corpus callosum (from brain midline to lateral ventricle, at bregma level −3), and internal capsule. (b) The expression of TLR2 in rat cortex, corpus callosum, and internal capsule was measured at 1 d, 3 d, and 7 d after DAI by immumohistochemical staining (scale bar = 100 μ m; n = 6). The bar graphs showed the statistical results for TLR2 expression ( ∗ p < 0.05, compared with control group; # p < 0.05, compared with DAI 1 d group; & p < 0.05, compared with DAI 3 d group). TLR2 positive cell levels were counted in 10 random fields (40x magnification). The relative increase in TLR2 level was assessed by densitometry (IOD). Data was presented as mean ± SD in three separate experiments.

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

Techniques: Expressing, Staining

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: Localization of TLR2 in rat cortex at 3 d after DAI. Double immunofluorescent staining was performed with antibody to TLR2 (red or green) and antibodies to NeuN (marker for neurons, green), GFAP (marker for astrocytes, green), and Iba-1 (marker for microglia, red), respectively (scale bar = 50 μ m; n = 6). The small figures located in the bottom right corner were the magnification of particular sections.

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

Techniques: Staining, Marker

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: Intracerebroventricular injection of TLR2 siRNA attenuated TLR2 expression. The feasibility and efficiency of TLR2 siRNA transfection were assessed. ((a), (b)) The normal brain was treated with TLR2 siRNA or control siRNA and cortexes were collected 1 d or 3 d later and subjected to western blot analysis and RT-PCR to measure TLR2 level, respectively. The expression of β -actin was used as an internal control. Values were presented as mean ± SD ( n = 6; ∗∗ p > 0.05, ∗ p < 0.05, compared with the control group; # p < 0.05, compared with the control siRNA group; & p < 0.05, compared with the 1 d postinfection group). (c) To confirm that TLR2 siRNA was effectively maintained in the DAI rats, immunofluorescence staining was carried out using antibodies to TLR2 (red) and DAPI (blue) in corpus callosum, respectively (scale bar = 100 μ m). The relative changes in TLR2 level were assessed by integral optical density (IOD). Values were presented as mean ± SD in three separate experiments ( n = 6; ∗ p < 0.05, compared with the control group; # p < 0.05, compared with the DAI 3 d + TLR2 siRNA group).

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

Techniques: Injection, Expressing, Transfection, Western Blot, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Staining

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: Role of TLR2 on secondary neuronal and axonal injury after DAI. (a) To confirm the role of TLR2 on the secondary neuronal injury, immumohistochemical staining of β -APP, tau, NF-L, and H&E staining in the rat cortex was performed (scale bar = 100 μ m). (b) The bar graphs showed the statistical results for β -APP, tau, and NF-L assessed by immunohistochemistry. The relative changes in β -APP, tau, and NF-L levels were counted in 10 random fields of relevant region (40x magnification) and assessed by densitometry (IOD). (c) The expression of β -APP and tau in each group was determined by western blotting. β -Actin was used as an internal control. (d) The bar graphs showed the statistical results for β -APP and tau expression. Data were presented as mean ± SD in three separate experiments ( n = 6; ∗ p < 0.05, compared with control group; # p < 0.05, compared with DAI 3 d group).

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

Techniques: Staining, Immunohistochemistry, Expressing, Western Blot

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: Role of TLR2 in ultrastructural alterations of axonal after DAI. Ultrastructural alterations of the longitudinal sections of axons after DAI were observed by TEM examination (scale bar = 5 μ m and 500 nm, resp.; n = 6).

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

Techniques:

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: TLR2 signaling was related to cell apoptosis after DAI. TUNEL assay was used to detect apoptotic cells in rat cortex. (a) Colocalization of TUNEL (green) and nuclei (DAPI, blue) were considered as apoptotic cells (scale bar = 100 μ m; enlarged figures scale bar = 20 μ m). (b) The bar graphs showed the statistical results of apoptotic cells accounts. The positive cell levels were counted in 10 random fields of relevant region (40x magnification). Data were presented as mean ± SD in three separate experiments ( n = 6; ∗ p < 0.05, compared with control group; # p < 0.05, compared with DAI 3 d group). Apoptotic index (AI), defined as the number of TUNEL positive cells per mm 2 .

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

Techniques: TUNEL Assay

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: Inhibition or activation of TLR2 was associated with glial reaction. (a) Marker of astrocyte vimentin, GFAP, and marker of microglial cells Iba-1 were assessed by immunohistochemistry in rat cortex (scale bar = 100 μ m). (b) The bar graphs showed the statistical results for vimentin, GFAP, and Iba-1 assessed by immunohistochemistry. The relative increases in vimentin, GFAP, and Iba-1 levels were assessed by densitometry (IOD) and positive cell levels were calculated in 10 random fields (40x magnification). Data were presented as mean ± SD in three separate experiments ( n = 6; ∗ p < 0.05, compared with control group; # p < 0.05, compared with DAI 3 d group).

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

Techniques: Inhibition, Activation Assay, Marker, Immunohistochemistry

Journal: Mediators of Inflammation

Article Title: Involvement of Toll Like Receptor 2 Signaling in Secondary Injury during Experimental Diffuse Axonal Injury in Rats

doi: 10.1155/2017/1570917

Figure Lengend Snippet: TLR2 signaling pathway induced-inflammatory factors were involved in secondary injury after DAI. (a) Phosphorylation of signaling molecules including p38 MAPK, ERK1/2, and JNK in cortex was examined by western blotting. The expression of β -actin was used as an internal control. (b) The bar graphs showed the statistical results for phosphorylation levels of p38 MAPK, ERK1/2, and JNK. (c) Expression of NF- κ B p65 and phospho-NF- κ B p65 in nuclear extraction of cortex was examined by western blotting. The expression of lamin B was used as an internal control. (d) The bar graphs showed the statistical results for NF- κ B p65 and phospho-NF- κ B p65 in nuclear extraction. (e) The effects of TLR2 siRNA and Pam3CSK4 on the levels of inflammatory factors including TNF- α , IL-1 β , and IL-6 in rat cortex after DAI were determined by ELISA. All data were presented as mean ± SD in three separate experiments ( n = 6; ∗ p < 0.05, compared with control group; # p < 0.05, compared with DAI 3 d group).

Article Snippet: Sections were then blocked in 2% (v/v) normal donkey serum and incubated in primary antibodies including rabbit polyclonal TLR2 (1 : 100, Bioss, Beijing, China), rabbit monoclonal β -APP (1 : 200, Abcam, Cambridge, UK), mouse monoclonal tau46 (1 : 300, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal neurofilament-L (NF-L, 1 : 100, Cell Signaling Technology, Danvers, MA, USA), mouse monoclonal GFAP (1 : 400, Cell Signaling Technology, Danvers, MA, USA), rabbit monoclonal vimentin (1 : 400, Cell Signaling Technology, Danvers, MA, USA), and rabbit monoclonal Iba-1 (1 : 400, Wako, Osaka, Japan) overnight at 4°C.

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

Journal: BMC Microbiology

Article Title: Advanced application of bovine intestinal epithelial cell line for evaluating regulatory effect of lactobacilli against heat-killed enterotoxigenic Escherichia coli -mediated inflammation

doi: 10.1186/1471-2180-13-54

Figure Lengend Snippet: Primer sequences used in this study

Article Snippet: Cells were then incubated with Alexa 488 conjugated rabbit anti-TLR2 polyclonal antibody (bs-1019R-Alexa488, Bioss Inc., Wobum, MA, USA) or Alexa 488 conjugated rabbit anti-TLR4 antibody (bs-1021R-Alexa488, Bioss) diluted 50 times with Can Get Signal solution 1 (NKB-201, TOYOBO Co., Ltd., Osaka, Japan) overnight at 4°C.

Techniques:

Journal: BMC Microbiology

Article Title: Advanced application of bovine intestinal epithelial cell line for evaluating regulatory effect of lactobacilli against heat-killed enterotoxigenic Escherichia coli -mediated inflammation

doi: 10.1186/1471-2180-13-54

Figure Lengend Snippet: Analysis of toll - like receptors (TLRs) expression in bovine intestinal epithelial (BIE) cells. ( A ) TLR1-10 mRNA levels in BIE cells. The expression of TLR in BIE cells was calculated first as relative units compared to bovine β-actin level. After calculating the relative unit to β-actin, TLR1 was set as 1. Values represent means and error bars indicate the standard deviations. The results are means of six independent experiments. ( B ) Immunofluorescent localization of TLR2 and TLR4 in BIE cells. Green images indicate bovine TLR2 or TLR4 positive cells and nuclei in all panels were stained with DAPI (blue). Control experiments were performed by omitting the primary antibody. The results represent six independent experiments.

Article Snippet: Cells were then incubated with Alexa 488 conjugated rabbit anti-TLR2 polyclonal antibody (bs-1019R-Alexa488, Bioss Inc., Wobum, MA, USA) or Alexa 488 conjugated rabbit anti-TLR4 antibody (bs-1021R-Alexa488, Bioss) diluted 50 times with Can Get Signal solution 1 (NKB-201, TOYOBO Co., Ltd., Osaka, Japan) overnight at 4°C.

Techniques: Expressing, Staining

Journal: PLoS ONE

Article Title: Stiffening-Induced High Pulsatility Flow Activates Endothelial Inflammation via a TLR2/NF-κB Pathway

doi: 10.1371/journal.pone.0102195

Figure Lengend Snippet: (A-B) High pulsatility flow (HPF), due to the use of a “stiff” tube upstream to cell culture, upregulated proinflammatory molecule mRNA (ICAM-1, VCAM-1, MCP-1 and E-selectin) and protein (MCP-1) in healthy PAECs, compared to low pulsatility flow (LPF) or to static conditions. (C-D) The mRNA and protein expression of TLR2 but not TLR4 in PAECs was highly upregulated by HPF. *: p<0.05 versus Static, †: p<0.05 versus LPF.

Article Snippet: Antibodies used here include: rabbit polyclonal antibody against bovine MCP-1 (1∶500 dilution; Kingfisher Biotech, St. Paul, MN), rabbit polyclonal antibody against TLR2 (1∶100 dilution; Bioss Inc, Woburn MA), mouse monoclonal antibody against TLR4 (1∶100 dilution; Acris Antibodies Inc, San Diego, CA) and mouse monoclonal antibody against GAPDH (1∶20,000 dilution; Sigma Inc, Saint Louis, MO).

Techniques: Cell Culture, Expressing

Journal: PLoS ONE

Article Title: Stiffening-Induced High Pulsatility Flow Activates Endothelial Inflammation via a TLR2/NF-κB Pathway

doi: 10.1371/journal.pone.0102195

Figure Lengend Snippet: (A, C) At the mRNA level, TLR2/4 inhibitor OxPAPC or TLR2 siRNA but not TLR4 inhibitor CLI-095, decreased PAEC expression of ICAM-1, VCAM-1, MCP-1 and E-selectin mRNAs under HPF; TLR4 siRNA decreased ICAM-1 and E-selectin but not VCAM-1 and MCP-1 mRNAs. “*”: p<0.05 versus LPF, “†”: p<0.05 versus HPF. (B, D) At the protein level, the MCP-1 expression in PAECs exposed to HPF was inhibited by OxPAPC or TLR2 siRNA treatment but not CLI-095 or TLR4 siRNA. The black line in the blot images (D, right) shows separated lanes obtained on the same gel.

Article Snippet: Antibodies used here include: rabbit polyclonal antibody against bovine MCP-1 (1∶500 dilution; Kingfisher Biotech, St. Paul, MN), rabbit polyclonal antibody against TLR2 (1∶100 dilution; Bioss Inc, Woburn MA), mouse monoclonal antibody against TLR4 (1∶100 dilution; Acris Antibodies Inc, San Diego, CA) and mouse monoclonal antibody against GAPDH (1∶20,000 dilution; Sigma Inc, Saint Louis, MO).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Stiffening-Induced High Pulsatility Flow Activates Endothelial Inflammation via a TLR2/NF-κB Pathway

doi: 10.1371/journal.pone.0102195

Figure Lengend Snippet: Pulse flow modulates TLR2-induced NF-κB activation in BPAECs. (A) Representative fluorescent images and quantitative measures of NF-kBp65 staining (red) in PAECs show HPF stimulation of PAECs led to increased intranuclear translocation or activation of NF-κB, which was reduced by TLR2/4 inhibitor OxPAPC and TLR2 siRNA. Blue stains show the nuclei. The scale bar shows 50 µm. (B) HPF stimulation of PAECs increased the mRNA levels of IKKα and IKKβ, both of which were attenuated in siRNA-transfected cells with knockdown of TLR2. (C) NF-κB inhibitor (BAY 11–7082) decreased the MCP-1 expression by PAECs exposed to HPF. *: p<0.05 versus LPF, †: p<0.05 versus HPF.

Article Snippet: Antibodies used here include: rabbit polyclonal antibody against bovine MCP-1 (1∶500 dilution; Kingfisher Biotech, St. Paul, MN), rabbit polyclonal antibody against TLR2 (1∶100 dilution; Bioss Inc, Woburn MA), mouse monoclonal antibody against TLR4 (1∶100 dilution; Acris Antibodies Inc, San Diego, CA) and mouse monoclonal antibody against GAPDH (1∶20,000 dilution; Sigma Inc, Saint Louis, MO).

Techniques: Activation Assay, Staining, Translocation Assay, Transfection, Expressing

Journal: PLoS ONE

Article Title: Stiffening-Induced High Pulsatility Flow Activates Endothelial Inflammation via a TLR2/NF-κB Pathway

doi: 10.1371/journal.pone.0102195

Figure Lengend Snippet: (A) PAECs from calves with hypoxia-induced pulmonary hypertension (PH) show elevated expression of TLR2 and TLR4, compared to control (CO). *:p<0.05. (B) Both immunostaining and western blotting results show elevated MCP-1 expression by PH-ECs compared to CO-ECs from calves. “PA” indicates the lumen of a pulmonary artery. *:p<0.05. (C) Enhanced TLR2 expression in the pulmonary arterial endothelium of human with pulmonary arterial hypertension (PAH). Cryosections of human intra-lobar pulmonary arteries were immunostained with TLR2 (red fluorescence) and counterstained with DAPI (cell nuclei, blue). Elastic lamellae showed green auto-fluorescence.

Article Snippet: Antibodies used here include: rabbit polyclonal antibody against bovine MCP-1 (1∶500 dilution; Kingfisher Biotech, St. Paul, MN), rabbit polyclonal antibody against TLR2 (1∶100 dilution; Bioss Inc, Woburn MA), mouse monoclonal antibody against TLR4 (1∶100 dilution; Acris Antibodies Inc, San Diego, CA) and mouse monoclonal antibody against GAPDH (1∶20,000 dilution; Sigma Inc, Saint Louis, MO).

Techniques: Expressing, Immunostaining, Western Blot, Fluorescence