the polyclonal antibody against cox-2  (Cayman Chemical)

Journal: Journal of Neuroinflammation

Article Title: Celecoxib reduces brain dopaminergic neuronaldysfunction, and improves sensorimotor behavioral performance in neonatal rats exposed to systemic lipopolysaccharide

doi: 10.1186/1742-2094-10-45

Figure Lengend Snippet: Representative photomicrographs of COX-2 immunostaining (A to I, SN) in the rat brain after LPS injection. Weak COX-2 positive staining was detectable in the brain sections of control brain ( A ). LPS injection increased the expression of inducible cyclooxygenase, as indicated by COX-2+ staining, in SN ( B , D and G , red) and striatum ( J ) of P6 rat brain. Celecoxib treatment attenuated the LPS-increased COX-2+ staining in the above areas ( C and J ). F is a merged image of D and E . Double-labeling (yellow) showed that many COX-2 positive cells ( F ) were GFAP + cells (astrocytes) ( E , green). I is a merged image of G and H . Double-labeling (yellow) also showed that some COX-2 positive cells ( I ) were TH + cells (dopaminergic neuron) ( H , green). The scale bar in A represents 50 μm for A to I . Quantitation of the percentage area of image that contained COX-2+ staining in the SN and striatum was performed as described in Methods. The results are expressed as the mean ± SEM of six animals in each group, and analyzed by one-way ANOVA. * P <0.05 represents a significant difference for the LPS + Vehicle group as compared with the Saline + Vehicle group. # P <0.05 represents a significant difference for the LPS + Celecoxib group as compared with the LPS + Vehicle group.

Article Snippet: Polyclonal rat antibodies against dopamine transporter (DAT) and polyclonal goat antibodies against COX-2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Immunostaining, Injection, Staining, Expressing, Labeling, Quantitation Assay

Journal: Mediators of Inflammation

Article Title: Diethylcarbamazine Attenuates the Development of Carrageenan-Induced Lung Injury in Mice

doi: 10.1155/2014/105120

Figure Lengend Snippet: Effect of DEC on immunohistochemical localization of COX-2 in lung tissue after pleurisy induced by carrageenan. (a) In tissue sections from the CAR group, positive labeling was detected on type II pneumocytes. (b) Treatment with DEC significantly reduced COX-2 staining in comparison to the CAR group, achieving levels similar to the sham group. (c) Densitometry analysis of immunohistochemistry photographs for COX-2 from lung tissues; figure is representative of at least 3 experiments performed on different experimental days; data expressed as mean ± S.E.M. from n = 5 mice for each group; ND: not detected; * P < 0.05 versus carrageenan; scale bar = 20 μ m.

Article Snippet: After blocking overnight at 4°C with 5% nonfat milk in TBS-T (Tris-buffered saline 0.1% plus 0.05% Tween 20, pH 7.4), the membranes were incubated at room temperature for 3 h with rabbit polyclonal antibody against COX-2 (1 : 1,000 dilution; ABCAM, CA, USA), IL-1 β (1 : 2,000 dilution, Genway, San Diego, CA, USA) and NF κ B (1 : 200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted in buffer solution TBS-T containing 3% nonfat milk.

Techniques: Immunohistochemical staining, Labeling, Staining, Immunohistochemistry

Journal: Mediators of Inflammation

Article Title: Diethylcarbamazine Attenuates the Development of Carrageenan-Induced Lung Injury in Mice

doi: 10.1155/2014/105120

Figure Lengend Snippet: Effects of DEC on carrageenan-induced COX-2 expression in the lung. Basal expression of COX-2 was detected in lung samples from the sham group, whereas COX-2 levels were substantially elevated in lung tissue obtained from animals 4 h after carrageenan injection. DEC treatment reduced the expression of COX-2, but treatment with indomethacin did not decrease the levels of COX-2 (a) and (b). (a) Representative blot of lysates obtained from pool 4 animals per group; (b) data expressed as mean ± S.E.M. of 4 replications for each group; * P < 0.05 versus carrageenan; ° P < 0.05 versus DEC.

Article Snippet: After blocking overnight at 4°C with 5% nonfat milk in TBS-T (Tris-buffered saline 0.1% plus 0.05% Tween 20, pH 7.4), the membranes were incubated at room temperature for 3 h with rabbit polyclonal antibody against COX-2 (1 : 1,000 dilution; ABCAM, CA, USA), IL-1 β (1 : 2,000 dilution, Genway, San Diego, CA, USA) and NF κ B (1 : 200 dilution, Santa Cruz Biotechnology, Santa Cruz, CA, USA) diluted in buffer solution TBS-T containing 3% nonfat milk.

Techniques: Expressing, Injection

Journal: Gene Regulation and Systems Biology

Article Title: Tetracycline Derivative Minocycline Inhibits Autophagy and Inflammation in Concanavalin-A-Activated Human Hepatoma Cells

doi: 10.4137/GRSB.S13946

Figure Lengend Snippet: Minocycline antagonizes the protein expression of BNIP3, MT1-MMP, and COX-2 in ConA-activated HepG2 cells. Serum-starved HepG2 hepatoma cells were treated with various ConA concentrations for 24 hours ( A ), or co-treated with ConA (30 μg/mL) having various concentrations of minocycline ( C ). Cell lysates were isolated and electrophoresed as described in the Methods sections. Scanning densitometry measurements were performed ( B, D ) on two independent experiments. A representative scanning densitometry profile is shown for the respective treatments. Data represent mean values in duplicates.

Article Snippet: The polyclonal antibody against COX-2 was from Cayman Chemical (Ann Arbor, MI).

Techniques: Expressing, Isolation

Journal: Gene Regulation and Systems Biology

Article Title: Tetracycline Derivative Minocycline Inhibits Autophagy and Inflammation in Concanavalin-A-Activated Human Hepatoma Cells

doi: 10.4137/GRSB.S13946

Figure Lengend Snippet: Minocycline exerts anti-inflammatory and anti-autophagy action in ConA-activated hepatoma cells. ConA triggers STAT3 phosphorylation and NANOS1 gene expression in HepG2 cells. NANOS1 and STAT3 phosphorylation is upstream prerequisite for ConA-mediated MT1-MMP gene expression, which, in turn, triggers the inflammatory biomarker COX-2 expression, as well as the autophagy biomarker BNIP3 and acidic vacuole formation. Minocycline can exert its effects by inhibiting ConA-induced STAT3 phosphorylation, or directly on either MT1-MMP or NANOS1 transcriptional regulation. Dotted lines : Minocycline exerts anti-angiogenic effects. An alternate feedback loop was recently suggested to occur, where MT1-MMP contributed to STAT3 phosphorylation, which in turn impacted on proangiogenic transcription of proangiogenic cytokines.

Article Snippet: The polyclonal antibody against COX-2 was from Cayman Chemical (Ann Arbor, MI).

Techniques: Expressing, Biomarker Assay

Journal: PLoS ONE

Article Title: Let-7b Is Involved in the Inflammation and Immune Responses Associated with Helicobacter pylori Infection by Targeting Toll-Like Receptor 4

doi: 10.1371/journal.pone.0056709

Figure Lengend Snippet: (A–B) H.pylori infection increased the expression of MyD88, p65/RelA, p50/NF-κB1 and IL-8 mRNA levels, and enhanced the NF-κB luciferase activity. (C–D) Overexpression of let-7b by mimics downregulated MyD88, p65/RelA, P50/NF-κB1 and IL-8 mRNA levels, and decreased NF-κB luciferase activity compared to let-7b negative control. Transfection with let-7b mimics inhibited NF-κB luciferase activity, irrespective of H.pylori infection. (E) Overexpression of let-7b by mimics decreased the expression of COX-2 and CyclinD1 in a dose-dependent manner. (F) Effects of let-7b on H.pylori -induced COX-2 and CyclinD1 expression in AGS cells were assessed by western blot. Data were expressed as mean ± S.D. from three independent experiments. *p<0.05, **p<0.01, ***p<0.001, # p<0.05.

Article Snippet: The blots were stripped using stripping buffer for 20 min at 60°C and reprobed using rabbit polyclonal antibody against human CyclinD1 (Santa Cruz, USA) at a dilution of 1∶300 and polyclonal antibody against COX-2 (Santa Cruz, USA) at a dilution of 1∶150.

Techniques: Infection, Expressing, Luciferase, Activity Assay, Over Expression, Negative Control, Transfection, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Anti-inflammatory and anti-amyloidogenic effects of a small molecule, 2,4-bis(p-hydroxyphenyl)-2-butenal in Tg2576 Alzheimer’s disease mice model

doi: 10.1186/1742-2094-10-2

Figure Lengend Snippet: HPB242 inhibited activation of astrocytes and microglia, and reduced expression of COX-2 and iNOS in the brains of Tg2576 mice. The effect of HPB242 on reactive astrocytes and activated microglia cells was measured by immunohistochemical analysis and western blotting analysis. The sections of mice brain incubated with anti-glial fibrillary acidic protein (GFAP) ( A ) or ionized calcium binding adaptor molecule 1 (Iba1) ( B ) primary antibody and the biotinylated secondary antibody (n = 8). The stained tissues were viewed with a microscope (×100 or 400). Expression of GFAP and Iba1 were also examined by specific antibodies in the cortex and hippocampus ( C ). Each blot is representative for three experiments (n = 8). Inhibitory effects of HPB242 on the Tg2576 mice brain expression of inflammatory proteins. The sections of mouse brain incubated with anti-iNOS ( D ) or COX-2 ( E ) and the biotinylated secondary antibody (n=8). The resulting tissue was viewed with a microscope (×100 or 400). The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were detected by western blotting using specific antibodies ( F ). Each blot is representative for three experiments (n = 8). *Significantly different to non-treated Tg2576 mice ( P < 0.05).

Article Snippet: Sections were then incubated for 2 h at room temperature with a mouse polyclonal antibody against Aβ (1:5000; Covance, Berkeley, CA, USA), a rabbit polyclonal antibody against GFAP and iNOS (1:1000; Abcam, Inc, Cambridge, MA, USA), a rabbit polyclonal antibody against COX-2 (1:1000; Cayman, Ann Arbor, MI, USA) and a rabbit polyclonal antibody against ionized calcium binding adaptor molecule 1 (Iba1) (1:1000; Wako, Osaka, Japan).

Techniques: Activation Assay, Expressing, Immunohistochemical staining, Western Blot, Incubation, Binding Assay, Staining, Microscopy