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Santa Cruz Biotechnology anti inos primer antibody
Photomicrographsof <t>nNOS,</t> <t>iNOS</t> and eNOS staining within the control and IBS tissues is depicted (×10). The immunoreactivity of sections stained with (a and b) nNOS (c and d) iNOS and (e and f) eNOS primary antibodies. Intensity of reactivity (score: 0= no staining; 1= mild; 2= moderate, and; 3= intense). nNOS: Neuronal nitric oxide synthase, iNOS: Inducible nitric oxide synthase, eNOS: Endothelial nitric oxide synthase
Anti Inos Primer Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 80/100, based on 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome"

Article Title: Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome

Journal: Indian Journal of Pharmacology

doi: 10.4103/0253-7613.186189

Photomicrographsof nNOS, iNOS and eNOS staining within the control and IBS tissues is depicted (×10). The immunoreactivity of sections stained with (a and b) nNOS (c and d) iNOS and (e and f) eNOS primary antibodies. Intensity of reactivity (score: 0= no staining; 1= mild; 2= moderate, and; 3= intense). nNOS: Neuronal nitric oxide synthase, iNOS: Inducible nitric oxide synthase, eNOS: Endothelial nitric oxide synthase
Figure Legend Snippet: Photomicrographsof nNOS, iNOS and eNOS staining within the control and IBS tissues is depicted (×10). The immunoreactivity of sections stained with (a and b) nNOS (c and d) iNOS and (e and f) eNOS primary antibodies. Intensity of reactivity (score: 0= no staining; 1= mild; 2= moderate, and; 3= intense). nNOS: Neuronal nitric oxide synthase, iNOS: Inducible nitric oxide synthase, eNOS: Endothelial nitric oxide synthase

Techniques Used: Staining

2) Product Images from "The role of Galectin-3 in α-synuclein-induced microglial activation"

Article Title: The role of Galectin-3 in α-synuclein-induced microglial activation

Journal: Acta Neuropathologica Communications

doi: 10.1186/s40478-014-0156-0

Galectin-3 siRNA reduces microglial activation induced by α-synuclein aggregates. BV2 microglia activated by 20 μM of α-synuclein aggregates for 12 h show a robust iNOS down regulation by 80% when galectin-3 is knocked down by siRNA (B) . Knock down efficiency of galectin-3 siRNA (A) . The cytokines levels from BV2 cells treated medium was measured after 12 h incubation with α-synuclein aggregates and we found significant reduction in TNF-α and IL-10 (C) . Western blot analysis showing iNOS and β-actin protein levels. t-test, One-Way ANOVA. *P
Figure Legend Snippet: Galectin-3 siRNA reduces microglial activation induced by α-synuclein aggregates. BV2 microglia activated by 20 μM of α-synuclein aggregates for 12 h show a robust iNOS down regulation by 80% when galectin-3 is knocked down by siRNA (B) . Knock down efficiency of galectin-3 siRNA (A) . The cytokines levels from BV2 cells treated medium was measured after 12 h incubation with α-synuclein aggregates and we found significant reduction in TNF-α and IL-10 (C) . Western blot analysis showing iNOS and β-actin protein levels. t-test, One-Way ANOVA. *P

Techniques Used: Activation Assay, Incubation, Western Blot

Inhibition of microglial activation by galectin-3 inhibitor. To determine the role of galectin-3 we used a treatment, incubating the galectin-3 inhibitor along with α-synuclein aggregates for 12 h at 20 μM. We determine by western blot the iNOS expression induced by α-synuclein aggregates. iNOS expression was inhibited by more than 80% using 100 μM of the inhibitor (A) . The cytokines levels were measure and TNF-α, IL-12 and IL-6 were down regulated when using the inhibitor for 12 along with α-synuclein aggregates (B) . We use the highest iNOS response in each experiment as an internal control to evaluate the response to the other concentrations. Western blot analysis displays iNOS and β-actin protein levels. One-way ANOVA, *P
Figure Legend Snippet: Inhibition of microglial activation by galectin-3 inhibitor. To determine the role of galectin-3 we used a treatment, incubating the galectin-3 inhibitor along with α-synuclein aggregates for 12 h at 20 μM. We determine by western blot the iNOS expression induced by α-synuclein aggregates. iNOS expression was inhibited by more than 80% using 100 μM of the inhibitor (A) . The cytokines levels were measure and TNF-α, IL-12 and IL-6 were down regulated when using the inhibitor for 12 along with α-synuclein aggregates (B) . We use the highest iNOS response in each experiment as an internal control to evaluate the response to the other concentrations. Western blot analysis displays iNOS and β-actin protein levels. One-way ANOVA, *P

Techniques Used: Inhibition, Activation Assay, Western Blot, Expressing

Microglial activation by α-synuclein and inhibition by galectin-3 inhibitor. We measured iNOS expression by western blot in microglial cells after 12 h incubation with α-synuclein monomers (A) and α-synuclein aggregates (B) using different concentrations, 5 μM, 10 μM and 20 μM. iNOS was significantly up regulated with both protein preparations of α-synuclein. α-synuclein aggregates (B) induced a 3-fold higher activation compared to monomers (A) . To determine the role of galectin-3 we used a pre-treatment, incubating the galectin-3 inhibitor for 30 min and then we incubated for 12 h the cells with α-synuclein, monomers or aggregates, using the highest concentration, 20 μM. The lower iNOS expression induced by α-synuclein monomers was not significantly inhibited by pharmacological inhibition of galectin-3 (C) . iNOS expression induced by α-synuclein aggregates (D) was inhibited by more than 50% using 100 μM of the inhibitor. We use the highest iNOS response in each experiment as an internal control to evaluate the response to the other concentrations. Western blot analysis displays iNOS and β-actin protein levels. One-way ANOVA, *P
Figure Legend Snippet: Microglial activation by α-synuclein and inhibition by galectin-3 inhibitor. We measured iNOS expression by western blot in microglial cells after 12 h incubation with α-synuclein monomers (A) and α-synuclein aggregates (B) using different concentrations, 5 μM, 10 μM and 20 μM. iNOS was significantly up regulated with both protein preparations of α-synuclein. α-synuclein aggregates (B) induced a 3-fold higher activation compared to monomers (A) . To determine the role of galectin-3 we used a pre-treatment, incubating the galectin-3 inhibitor for 30 min and then we incubated for 12 h the cells with α-synuclein, monomers or aggregates, using the highest concentration, 20 μM. The lower iNOS expression induced by α-synuclein monomers was not significantly inhibited by pharmacological inhibition of galectin-3 (C) . iNOS expression induced by α-synuclein aggregates (D) was inhibited by more than 50% using 100 μM of the inhibitor. We use the highest iNOS response in each experiment as an internal control to evaluate the response to the other concentrations. Western blot analysis displays iNOS and β-actin protein levels. One-way ANOVA, *P

Techniques Used: Activation Assay, Inhibition, Expressing, Western Blot, Incubation, Concentration Assay

Abrogation of iNOS proteins level and pro-inflammatory cytokines reduction in primary microglial cells from galectin-3 knockout mice after activation with α-synuclein. Primary microglial culture from wild-type mice shows robust iNOS expression following exposure of 20 μM α-synuclein aggregates, or LPS (100 ng/ml), for 12 h (A) . Lower concentrations of α-synuclein aggregates, 5 μM and below, failed to induce iNOS expression in wild- type microglia (A) . Primary microglia from galectin-3 knockout mice completely lack iNOS up regulation following exposure of 20 μM α-synuclein aggregates for 12 h (B) . Cytokine levels in culture medium from primary microglial cells were measured after 12 h incubation with α-synuclein aggregates. Treatment of wild-type microglia with 5 and 20 μM α-synuclein aggregates for 12 h induced increased levels of IL-1β, IL-12, IFN-γ and IL-4 (C) . Treatment of galectin-3 knockout microglia for 12 h reduced levels of IL-1β IL-12 using 20 μM α-synuclein aggregates. Cytokine levels of IFN-γ and IL-4 did not change in galectin-3 knockout compared to wild-type microglia. Two-way ANOVA, *P
Figure Legend Snippet: Abrogation of iNOS proteins level and pro-inflammatory cytokines reduction in primary microglial cells from galectin-3 knockout mice after activation with α-synuclein. Primary microglial culture from wild-type mice shows robust iNOS expression following exposure of 20 μM α-synuclein aggregates, or LPS (100 ng/ml), for 12 h (A) . Lower concentrations of α-synuclein aggregates, 5 μM and below, failed to induce iNOS expression in wild- type microglia (A) . Primary microglia from galectin-3 knockout mice completely lack iNOS up regulation following exposure of 20 μM α-synuclein aggregates for 12 h (B) . Cytokine levels in culture medium from primary microglial cells were measured after 12 h incubation with α-synuclein aggregates. Treatment of wild-type microglia with 5 and 20 μM α-synuclein aggregates for 12 h induced increased levels of IL-1β, IL-12, IFN-γ and IL-4 (C) . Treatment of galectin-3 knockout microglia for 12 h reduced levels of IL-1β IL-12 using 20 μM α-synuclein aggregates. Cytokine levels of IFN-γ and IL-4 did not change in galectin-3 knockout compared to wild-type microglia. Two-way ANOVA, *P

Techniques Used: Knock-Out, Mouse Assay, Activation Assay, Expressing, Incubation

3) Product Images from "The composition of a bioprocessed shiitake (Lentinus edodes) mushroom mycelia and rice bran formulation and its antimicrobial effects against Salmonella enterica subsp. enterica serovar Typhimurium strain SL1344 in macrophage cells and in mice"

Article Title: The composition of a bioprocessed shiitake (Lentinus edodes) mushroom mycelia and rice bran formulation and its antimicrobial effects against Salmonella enterica subsp. enterica serovar Typhimurium strain SL1344 in macrophage cells and in mice

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/s12906-018-2365-8

Effects of BPRBE on iNOS gene expression in S. Typhimurium-infected macrophages. a iNOS mRNA expression profiles assessed by RT-PCR. RAW 264.7 cells were cultivated with or without BPRBE for 24 h. The cells were then infected with S. Typhimurium SL1344 for 8 h. After infection, total RNA was purified and an iNOS mRNA expression profile was determined using RT-PCR analysis. b Assessment of iNOS protein expression profiles by Western blot analysis. RAW 264.7 cells treated with BPRBE for 24 h and then infected with S. Typhimurium were lysed and the iNOS protein in the cell lysate was then identified by Western blotting using rabbit anti-mouse iNOS polyclonal antibody. The relative proportions of iNOS mRNA and polypeptide are expressed as the R. E. (relative expression) values calculated from iNOS/β-actin gene expressions. Figures represent results from at least three individual experiments
Figure Legend Snippet: Effects of BPRBE on iNOS gene expression in S. Typhimurium-infected macrophages. a iNOS mRNA expression profiles assessed by RT-PCR. RAW 264.7 cells were cultivated with or without BPRBE for 24 h. The cells were then infected with S. Typhimurium SL1344 for 8 h. After infection, total RNA was purified and an iNOS mRNA expression profile was determined using RT-PCR analysis. b Assessment of iNOS protein expression profiles by Western blot analysis. RAW 264.7 cells treated with BPRBE for 24 h and then infected with S. Typhimurium were lysed and the iNOS protein in the cell lysate was then identified by Western blotting using rabbit anti-mouse iNOS polyclonal antibody. The relative proportions of iNOS mRNA and polypeptide are expressed as the R. E. (relative expression) values calculated from iNOS/β-actin gene expressions. Figures represent results from at least three individual experiments

Techniques Used: Expressing, Infection, Reverse Transcription Polymerase Chain Reaction, Purification, Western Blot

4) Product Images from "Evaluation of Antioxidant, Anti-cholinesterase, and Anti-inflammatory Effects of Culinary Mushroom Pleurotus pulmonarius"

Article Title: Evaluation of Antioxidant, Anti-cholinesterase, and Anti-inflammatory Effects of Culinary Mushroom Pleurotus pulmonarius

Journal: Mycobiology

doi: 10.5941/MYCO.2016.44.4.291

Inhibitory effect of Pleurotus pulmunarius methanol extract on lipopolysaccharide (LPS)-induced nitric oxide production and expression of inducible nitric oxide synthase (iNOS) in RAW 264.7 cells. A, Nitric oxide production; B, Expression of iNOS. β-Actin was used as an internal control. Accumulated nitric oxide in the culture medium was determined by the Griess method. The values are means ± SD (n = 3). *** p ≤ 0.001 vs. LPS treated group.
Figure Legend Snippet: Inhibitory effect of Pleurotus pulmunarius methanol extract on lipopolysaccharide (LPS)-induced nitric oxide production and expression of inducible nitric oxide synthase (iNOS) in RAW 264.7 cells. A, Nitric oxide production; B, Expression of iNOS. β-Actin was used as an internal control. Accumulated nitric oxide in the culture medium was determined by the Griess method. The values are means ± SD (n = 3). *** p ≤ 0.001 vs. LPS treated group.

Techniques Used: Expressing

5) Product Images from "Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes"

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes

Journal: Neuroscience

doi: 10.1016/j.neuroscience.2010.09.025

Laser (632.8nm) suppressed Aβ-induced expressions of IL-1β and iNOS in astrocytes
Figure Legend Snippet: Laser (632.8nm) suppressed Aβ-induced expressions of IL-1β and iNOS in astrocytes

Techniques Used:

6) Product Images from "Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes"

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2018/9125601

Effects of DOX and dl-NBP on neuroinflammation biomarkers: gene expression of IL-1 β (a), IL-6 (b), and TNF- α (c); protein expression of I κ B (e), p65 (f), and iNOS (g); and immunohistochemical staining of iNOS (h). Data are expressed as means ± SEM ( n = 8). ∗ p
Figure Legend Snippet: Effects of DOX and dl-NBP on neuroinflammation biomarkers: gene expression of IL-1 β (a), IL-6 (b), and TNF- α (c); protein expression of I κ B (e), p65 (f), and iNOS (g); and immunohistochemical staining of iNOS (h). Data are expressed as means ± SEM ( n = 8). ∗ p

Techniques Used: Expressing, Immunohistochemistry, Staining

7) Product Images from "The composition of a bioprocessed shiitake (Lentinus edodes) mushroom mycelia and rice bran formulation and its antimicrobial effects against Salmonella enterica subsp. enterica serovar Typhimurium strain SL1344 in macrophage cells and in mice"

Article Title: The composition of a bioprocessed shiitake (Lentinus edodes) mushroom mycelia and rice bran formulation and its antimicrobial effects against Salmonella enterica subsp. enterica serovar Typhimurium strain SL1344 in macrophage cells and in mice

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/s12906-018-2365-8

Effects of BPRBE on iNOS gene expression in S. Typhimurium-infected macrophages. a iNOS mRNA expression profiles assessed by RT-PCR. RAW 264.7 cells were cultivated with or without BPRBE for 24 h. The cells were then infected with S. Typhimurium SL1344 for 8 h. After infection, total RNA was purified and an iNOS mRNA expression profile was determined using RT-PCR analysis. b Assessment of iNOS protein expression profiles by Western blot analysis. RAW 264.7 cells treated with BPRBE for 24 h and then infected with S. Typhimurium were lysed and the iNOS protein in the cell lysate was then identified by Western blotting using rabbit anti-mouse iNOS polyclonal antibody. The relative proportions of iNOS mRNA and polypeptide are expressed as the R. E. (relative expression) values calculated from iNOS/β-actin gene expressions. Figures represent results from at least three individual experiments
Figure Legend Snippet: Effects of BPRBE on iNOS gene expression in S. Typhimurium-infected macrophages. a iNOS mRNA expression profiles assessed by RT-PCR. RAW 264.7 cells were cultivated with or without BPRBE for 24 h. The cells were then infected with S. Typhimurium SL1344 for 8 h. After infection, total RNA was purified and an iNOS mRNA expression profile was determined using RT-PCR analysis. b Assessment of iNOS protein expression profiles by Western blot analysis. RAW 264.7 cells treated with BPRBE for 24 h and then infected with S. Typhimurium were lysed and the iNOS protein in the cell lysate was then identified by Western blotting using rabbit anti-mouse iNOS polyclonal antibody. The relative proportions of iNOS mRNA and polypeptide are expressed as the R. E. (relative expression) values calculated from iNOS/β-actin gene expressions. Figures represent results from at least three individual experiments

Techniques Used: Expressing, Infection, Reverse Transcription Polymerase Chain Reaction, Purification, Western Blot

8) Product Images from "Green Tea Extract Ameliorates Learning and Memory Deficits in Ischemic Rats via Its Active Component Polyphenol Epigallocatechin-3-gallate by Modulation of Oxidative Stress and Neuroinflammation"

Article Title: Green Tea Extract Ameliorates Learning and Memory Deficits in Ischemic Rats via Its Active Component Polyphenol Epigallocatechin-3-gallate by Modulation of Oxidative Stress and Neuroinflammation

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2012/163106

Effects of EGCG (2, 10, and 25 μ M) on expression of COX-2 and iNOS in BV-2 cells treated with lipopolysaccharide (LPS, 0.5 μ g/mL) for 24 h. Cultures were pretreated with EGCG for 1 h before the addition of LPS treatment. Bars represent the mean ± SE from three independent experiments. Densitometry analyses are presented as the relative ratio of protein/ β -actin protein and are represented as percentages of the LPS only group. *** P
Figure Legend Snippet: Effects of EGCG (2, 10, and 25 μ M) on expression of COX-2 and iNOS in BV-2 cells treated with lipopolysaccharide (LPS, 0.5 μ g/mL) for 24 h. Cultures were pretreated with EGCG for 1 h before the addition of LPS treatment. Bars represent the mean ± SE from three independent experiments. Densitometry analyses are presented as the relative ratio of protein/ β -actin protein and are represented as percentages of the LPS only group. *** P

Techniques Used: Expressing

9) Product Images from "Effects of Hypericum Perforatum, in a rodent model of periodontitis"

Article Title: Effects of Hypericum Perforatum, in a rodent model of periodontitis

Journal: BMC Complementary and Alternative Medicine

doi: 10.1186/1472-6882-10-73

A significant increase in iNOS expression, assayed by Western blot analysis, was detected in the tissue from ligature-treated rats (a, a1) . The treatment with Hypericum significantly reduced iNOS expression in the gingivomucosal tissues (a, a1). Moreover positive staining for iNOS (b see densitometry analysis f), and nitrotyrosine (d see densitometry analysis f) was observed in gingivomucosal tissue after ligature. In gingivomucosal tissue of Hypericum treated rats no positive staining was observed for iNOS (c see densitometry analysis f), nitrotyrosine (e, see densitometry analysis f). Figure is representative of at least 3 experiments performed on different experimental days. Densitometry data are expressed as % of total tissue area. Data are means of mean ± s.e.m. from N = 10 rats for each group. *P
Figure Legend Snippet: A significant increase in iNOS expression, assayed by Western blot analysis, was detected in the tissue from ligature-treated rats (a, a1) . The treatment with Hypericum significantly reduced iNOS expression in the gingivomucosal tissues (a, a1). Moreover positive staining for iNOS (b see densitometry analysis f), and nitrotyrosine (d see densitometry analysis f) was observed in gingivomucosal tissue after ligature. In gingivomucosal tissue of Hypericum treated rats no positive staining was observed for iNOS (c see densitometry analysis f), nitrotyrosine (e, see densitometry analysis f). Figure is representative of at least 3 experiments performed on different experimental days. Densitometry data are expressed as % of total tissue area. Data are means of mean ± s.e.m. from N = 10 rats for each group. *P

Techniques Used: Expressing, Western Blot, Staining

10) Product Images from "Simvastatin Attenuates the Oxidative Stress, Endothelial Thrombogenicity and the Inducibility of Atrial Fibrillation in a Rat Model of Ischemic Heart Failure"

Article Title: Simvastatin Attenuates the Oxidative Stress, Endothelial Thrombogenicity and the Inducibility of Atrial Fibrillation in a Rat Model of Ischemic Heart Failure

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms150814803

( A ) Immunohistochemical staining showed that eNOS expression in the MI group was lower than the sham, and eNOS expression in the MI + simvastatin group was higher than the MI group (endothelial line, red arrows); and ( B ) iNOS expression in the MI group was higher than the sham, and iNOS expression in the MI + simvastatin group was lower than the MI group (cytoplasmic brown area, black arrows).
Figure Legend Snippet: ( A ) Immunohistochemical staining showed that eNOS expression in the MI group was lower than the sham, and eNOS expression in the MI + simvastatin group was higher than the MI group (endothelial line, red arrows); and ( B ) iNOS expression in the MI group was higher than the sham, and iNOS expression in the MI + simvastatin group was lower than the MI group (cytoplasmic brown area, black arrows).

Techniques Used: Immunohistochemistry, Staining, Expressing

Alterations in eNOS, iNOS, SERCA, NCX ( A ), TFPI, TM, tPA ( B ) and Rac 1 ( C ) protein expression in the left atrium (eNOS, endothelial nitric oxide synthase; iNOS, induced nitric oxide synthase; NCX, sodium calcium exchanger; SERCA, sarcoplasmic reticulum calcium ATPase; TM, thrombomodulin; TFPI, tissue factor pathway inhibitor; tPA, tissue plasminogen activator; Rac 1, Rac 1 GTPase).
Figure Legend Snippet: Alterations in eNOS, iNOS, SERCA, NCX ( A ), TFPI, TM, tPA ( B ) and Rac 1 ( C ) protein expression in the left atrium (eNOS, endothelial nitric oxide synthase; iNOS, induced nitric oxide synthase; NCX, sodium calcium exchanger; SERCA, sarcoplasmic reticulum calcium ATPase; TM, thrombomodulin; TFPI, tissue factor pathway inhibitor; tPA, tissue plasminogen activator; Rac 1, Rac 1 GTPase).

Techniques Used: Expressing

11) Product Images from "Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes"

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes

Journal: Oxidative Medicine and Cellular Longevity

doi: 10.1155/2018/9125601

Effects of DOX and dl-NBP on neuroinflammation biomarkers: gene expression of IL-1 β (a), IL-6 (b), and TNF- α (c); protein expression of I κ B (e), p65 (f), and iNOS (g); and immunohistochemical staining of iNOS (h). Data are expressed as means ± SEM ( n = 8). ∗ p
Figure Legend Snippet: Effects of DOX and dl-NBP on neuroinflammation biomarkers: gene expression of IL-1 β (a), IL-6 (b), and TNF- α (c); protein expression of I κ B (e), p65 (f), and iNOS (g); and immunohistochemical staining of iNOS (h). Data are expressed as means ± SEM ( n = 8). ∗ p

Techniques Used: Expressing, Immunohistochemistry, Staining

12) Product Images from "Microglia P2Y6 receptor is related to Parkinson’s disease through neuroinflammatory process"

Article Title: Microglia P2Y6 receptor is related to Parkinson’s disease through neuroinflammatory process

Journal: Journal of Neuroinflammation

doi: 10.1186/s12974-017-0795-8

LPS-activated microglia and upregulated P2Y6R expression. a P2Y6R expression in BV-2 cells after LPS stimulation according to western blot. b iNOS and COX-2 levels after LPS stimulation according to western blot. c Inflammatory cytokine (TNF-α, iNOS, IL-6, COX-2, and MIP-2) and P2Y6R expression levels after LPS stimulation according to RT-PCR. The band intensity was quantified using studio lite imager and is presented relative to the level of β-actin. Data are shown as the mean ± SD of three independent experiments. * p
Figure Legend Snippet: LPS-activated microglia and upregulated P2Y6R expression. a P2Y6R expression in BV-2 cells after LPS stimulation according to western blot. b iNOS and COX-2 levels after LPS stimulation according to western blot. c Inflammatory cytokine (TNF-α, iNOS, IL-6, COX-2, and MIP-2) and P2Y6R expression levels after LPS stimulation according to RT-PCR. The band intensity was quantified using studio lite imager and is presented relative to the level of β-actin. Data are shown as the mean ± SD of three independent experiments. * p

Techniques Used: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

Knockdown of P2Y6R reduced the production of inflammatory cytokines. a RT-PCR and western blot analyses of P2Y6R from BV-2 cells transfected with P2Y6 siRNA. b Representative western blots showing iNOS and COX-2 levels from LPS-stimulated BV-2 cells transfected with P2Y6 siRNA. c Representative RT-PCR showing the expression levels of inflammatory cytokines (TNF-α, iNOS, IL-6, COX-2, and MIP-2) from LPS-stimulated BV-2 cells transfected with P2Y6R siRNA. The band intensity was quantified using studio lite imager and is presented relative to the level of β-actin. Data are shown as the mean ± SD of three independent experiments. * p
Figure Legend Snippet: Knockdown of P2Y6R reduced the production of inflammatory cytokines. a RT-PCR and western blot analyses of P2Y6R from BV-2 cells transfected with P2Y6 siRNA. b Representative western blots showing iNOS and COX-2 levels from LPS-stimulated BV-2 cells transfected with P2Y6 siRNA. c Representative RT-PCR showing the expression levels of inflammatory cytokines (TNF-α, iNOS, IL-6, COX-2, and MIP-2) from LPS-stimulated BV-2 cells transfected with P2Y6R siRNA. The band intensity was quantified using studio lite imager and is presented relative to the level of β-actin. Data are shown as the mean ± SD of three independent experiments. * p

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Expressing

UDP/P2Y6R signaling involved in the activation of microglia cells and the production of inflammatory cytokines. a The standard curve and the UDP concentration in the LPS-treated and control supernatants of BV-2 cells based on HPLC. b mRNA levels of MIP-2 in LPS, LPS+Apy- and LPS+MRS-treated BV-2 cells. c ELISA of MIP-2 secretion. d Morphological changes in LPS-stimulated and Apy/MRS pretreated primary microglia cells. Representative immunostained images for primary microglia cells (Iba-1, red ; DAPI, blue ). Scale bar = 100 μm. e Representative RT-PCR showing the expression levels of inflammatory cytokines (TNF-α, iNOS, IL-6, COX-2, and MIP-2) from LPS-stimulated primary microglia cells pretreated with Apy/MRS. The band intensity was quantified using studio lite imager and is presented relative to the level of β-actin. Data are shown as the mean ± SD of three independent experiments. * p
Figure Legend Snippet: UDP/P2Y6R signaling involved in the activation of microglia cells and the production of inflammatory cytokines. a The standard curve and the UDP concentration in the LPS-treated and control supernatants of BV-2 cells based on HPLC. b mRNA levels of MIP-2 in LPS, LPS+Apy- and LPS+MRS-treated BV-2 cells. c ELISA of MIP-2 secretion. d Morphological changes in LPS-stimulated and Apy/MRS pretreated primary microglia cells. Representative immunostained images for primary microglia cells (Iba-1, red ; DAPI, blue ). Scale bar = 100 μm. e Representative RT-PCR showing the expression levels of inflammatory cytokines (TNF-α, iNOS, IL-6, COX-2, and MIP-2) from LPS-stimulated primary microglia cells pretreated with Apy/MRS. The band intensity was quantified using studio lite imager and is presented relative to the level of β-actin. Data are shown as the mean ± SD of three independent experiments. * p

Techniques Used: Activation Assay, Concentration Assay, High Performance Liquid Chromatography, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Expressing

13) Product Images from "Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles"

Article Title: Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles

Journal: Nucleic Acid Therapeutics

doi: 10.1089/nat.2012.0390

cSCK-pa 100 mediated silencing of iNOS expression by siRNA. Anti-iNOS siRNA481 (100 nM) was transfected into mouse monocyte-macrophage RAW264.7 cells using PAEA 128 - b -PS 40 cSCK-pa 100 at different N/P ratios (for N/P=1, cSCK 0.62 μg/mL),
Figure Legend Snippet: cSCK-pa 100 mediated silencing of iNOS expression by siRNA. Anti-iNOS siRNA481 (100 nM) was transfected into mouse monocyte-macrophage RAW264.7 cells using PAEA 128 - b -PS 40 cSCK-pa 100 at different N/P ratios (for N/P=1, cSCK 0.62 μg/mL),

Techniques Used: Expressing, Transfection

iNOS mRNA silencing by cSCK•siRNA•GALA nanocomplex in RAW264.7 cells. (A) cytoxicity of PAEA 160 -b-PS 30 cSCK-pa 100 after 24 h in RAW cells that had been induced for 24 hours with LPS/γ-IFN as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
Figure Legend Snippet: iNOS mRNA silencing by cSCK•siRNA•GALA nanocomplex in RAW264.7 cells. (A) cytoxicity of PAEA 160 -b-PS 30 cSCK-pa 100 after 24 h in RAW cells that had been induced for 24 hours with LPS/γ-IFN as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium

Techniques Used:

Endocytosis inhibition and tracking studies. (A) Inhibition experiments. iNOS-induced RAW 264.7 cells were incubated with cytochalasin D (Cyto-D) (average of 2–50 μM), chlorpromazine (CPZ) (20 μM) and methyl-beta-cyclodextrin,
Figure Legend Snippet: Endocytosis inhibition and tracking studies. (A) Inhibition experiments. iNOS-induced RAW 264.7 cells were incubated with cytochalasin D (Cyto-D) (average of 2–50 μM), chlorpromazine (CPZ) (20 μM) and methyl-beta-cyclodextrin,

Techniques Used: Inhibition, Incubation

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Griess Assay:

Article Title: Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles
Article Snippet: The separated proteins were then transferred onto polyvinylidene difluoride membrane (Amersham Hybond™ -P, GE Healthcare). iNOS was detected by using monoclonal anti-iNOS antibody NOS2(C-11) at 1:200 (Santa Cruz Biotechnology, Inc.). β-actin was detected using mouse anti- β-actin monoclonal antibody at 1:20,000 (GenScript Corporation). .. The separated proteins were then transferred onto polyvinylidene difluoride membrane (Amersham Hybond™ -P, GE Healthcare). iNOS was detected by using monoclonal anti-iNOS antibody NOS2(C-11) at 1:200 (Santa Cruz Biotechnology, Inc.). β-actin was detected using mouse anti- β-actin monoclonal antibody at 1:20,000 (GenScript Corporation).

Blocking Assay:

Article Title: Simvastatin Attenuates the Oxidative Stress, Endothelial Thrombogenicity and the Inducibility of Atrial Fibrillation in a Rat Model of Ischemic Heart Failure
Article Snippet: Serial sections (6 μm) were cut from each paraffin block. .. The sections were incubated with anti-eNOS antibody (Santa Cruz, CA, USA) or anti-iNOS antibody (Santa Cruz, CA, USA) at a 1:50 dilution in PBS.

Article Title: Significance of vascular endothelial growth factor expression and its correlation with inducible nitric oxide synthase in gastric cancer
Article Snippet: Endogenous peroxidase was blocked by incubation of samples in hydrogen peroxide blocking reagent (Maixin Bio, Fuzhou). .. The anti-VEGF antibody is a polyclonal mouse anti-serum against VEGF of human origin and recognizes an amino-terminal epitope found in VEGF121,165,189 , and the anti-iNOS antibody is a monoclonal mouse anti-serum against the C-terminal domain of iNOS of human origin (The two antibodies were from Santa Cruz Biotechnology, Inc., CA).

Electrophoresis:

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes
Article Snippet: Equivalent amounts of protein from each sample (e.g., 40 μg) were diluted with Laemmli buffer, boiled for 5 min, subjected to electrophoresis in 7.5% SDS-polyacrylamide gels, and transferred to nitrocellulose membranes. .. Membranes were blocked for 1 h with 5% (w/v) nonfat dry milk in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) and were incubated overnight at 4°C in 3% (w/v) BSA with 0.02% (w/v) sodium azide in TBST with p-cPLA2 or cPLA2 antibodies (1:1000 dilution; Cell Signaling Technology, Beverly, MA), anti-IL-1β antibody (1:1000 dilution; R & D Systems, Minneapolis, MN), anti-iNOS antibody (1:1000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin antibody (Sigma, St. Louis, MO).

Incubation:

Article Title: The Traditional Herbal Medicine, Dangkwisoo-San, Prevents Cerebral Ischemic Injury through Nitric Oxide-Dependent Mechanisms
Article Snippet: .. Immunoblot analysis was performed with anti-eNOS and anti-phospho-eNOS (pS1177) antibodies (BD Biosciences, San Jose, CA), anti-Akt and anti-phospho-Akt (Ser 473) antibodies (Cell signaling, Danvers, MA), anti-nNOS and anti-iNOS antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) followed by incubation with secondary antibody conjugated with horseradish peroxidase. .. The intensity of chemiluminescence was measured using an ImageQuant LAS 4000 apparatus (GE Healthcare Life Sciences, Uppsala, Sweden).

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: .. Immunohistochemical Staining For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v /v ). .. Sections were then washed with PBS and incubated with secondary antibodies.

Article Title: Simvastatin Attenuates the Oxidative Stress, Endothelial Thrombogenicity and the Inducibility of Atrial Fibrillation in a Rat Model of Ischemic Heart Failure
Article Snippet: .. The sections were incubated with anti-eNOS antibody (Santa Cruz, CA, USA) or anti-iNOS antibody (Santa Cruz, CA, USA) at a 1:50 dilution in PBS. .. After washing with PBS twice for 5 min each time, incubation was performed for 1 h at room temperature with biotinylated rabbit anti-rat IgG (1:50) in PBS.

Article Title: Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome
Article Snippet: .. Then, sections were incubated with anti-nNOS primer antibody (sc-55521, Santa Cruz Biotechnology, Inc.), anti-iNOS primer antibody (sc-649, Santa Cruz Biotechnology, Inc.), anti-eNOS antibody (sc-654, Santa Cruz Biotechnology, Inc.) in a 1/100 dilution for 18 h at +4°C. .. They were then given an additional three 5-washes in PBS, followed by incubation with biotinylated IgG and administration of streptavidin peroxidase (Histostain Plus Kit Cat. No: 85-9043, Invitrogen).

Article Title: TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection
Article Snippet: .. Samples were Tissue slides were then incubated overnight at 4°C with a biotin anti-mouse F4/80 antibody (1:150, Abcam) and a polyclonal rabbit anti-mouse iNOS antibody (1:200, Santa Cruz). ..

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: .. For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v / v ). .. Sections were then washed with PBS and incubated with secondary antibodies.

Article Title: Significance of vascular endothelial growth factor expression and its correlation with inducible nitric oxide synthase in gastric cancer
Article Snippet: After washed with phosphate-buffered saline solution, the samples were incubated for 60 min with the primary antibodies. .. The anti-VEGF antibody is a polyclonal mouse anti-serum against VEGF of human origin and recognizes an amino-terminal epitope found in VEGF121,165,189 , and the anti-iNOS antibody is a monoclonal mouse anti-serum against the C-terminal domain of iNOS of human origin (The two antibodies were from Santa Cruz Biotechnology, Inc., CA).

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes
Article Snippet: .. Membranes were blocked for 1 h with 5% (w/v) nonfat dry milk in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) and were incubated overnight at 4°C in 3% (w/v) BSA with 0.02% (w/v) sodium azide in TBST with p-cPLA2 or cPLA2 antibodies (1:1000 dilution; Cell Signaling Technology, Beverly, MA), anti-IL-1β antibody (1:1000 dilution; R & D Systems, Minneapolis, MN), anti-iNOS antibody (1:1000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin antibody (Sigma, St. Louis, MO). .. Membranes were washed three times during a 15-min period with TBST and incubated with horseradish peroxidase-conjugated goat anti-mouse or anti-rabbit IgG antibody (1: 5000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) in 5% (w/v) nonfat dry milk in TBST at room temperature for 1 h. After washing with TBST for three times, the membrane was subjected to SuperSignal West Pico Chemiluminescent detection reagents from Pierce (Rockford, IL) to visualize bands.

Stripping Membranes:

Article Title: TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection
Article Snippet: Histology and immunofluorescence A longitudinal strip from the greater curvature of the stomach was excised and placed in 10% normal buffered formalin for 24 h, embedded in paraffin and processed routinely for hematoxylin and eosin (H & E) or immunofluorescence staining. .. Samples were Tissue slides were then incubated overnight at 4°C with a biotin anti-mouse F4/80 antibody (1:150, Abcam) and a polyclonal rabbit anti-mouse iNOS antibody (1:200, Santa Cruz).

Activity Assay:

Article Title: Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome
Article Snippet: The tissues were then treated with 2% trypsin (ab970, Abcam, Cambridge, UK) at 37°C for 15 min and incubated in 3% H2 O2 solution for 15 min to inhibit endogenous peroxidase activity. .. Then, sections were incubated with anti-nNOS primer antibody (sc-55521, Santa Cruz Biotechnology, Inc.), anti-iNOS primer antibody (sc-649, Santa Cruz Biotechnology, Inc.), anti-eNOS antibody (sc-654, Santa Cruz Biotechnology, Inc.) in a 1/100 dilution for 18 h at +4°C.

Expressing:

Article Title: TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection
Article Snippet: Double-labeled immunofluorescence was carried out to localize F4/80 and iNOS expression in murine stomach. .. Samples were Tissue slides were then incubated overnight at 4°C with a biotin anti-mouse F4/80 antibody (1:150, Abcam) and a polyclonal rabbit anti-mouse iNOS antibody (1:200, Santa Cruz).

Article Title: Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles
Article Snippet: Paragraph title: Inhibition of iNOS expression by cSCKs•siRNA complexes ... The separated proteins were then transferred onto polyvinylidene difluoride membrane (Amersham Hybond™ -P, GE Healthcare). iNOS was detected by using monoclonal anti-iNOS antibody NOS2(C-11) at 1:200 (Santa Cruz Biotechnology, Inc.). β-actin was detected using mouse anti- β-actin monoclonal antibody at 1:20,000 (GenScript Corporation).

BIA-KA:

Article Title: Green Tea Extract Ameliorates Learning and Memory Deficits in Ischemic Rats via Its Active Component Polyphenol Epigallocatechin-3-gallate by Modulation of Oxidative Stress and Neuroinflammation
Article Snippet: BCA Protein assay kit was purchased from Thermo Fisher Scientific (Lafayette, CO, USA). .. Anti-iNOS antibody (rabbit polyclonal to iNOS, sc-651) and anti-COX-2 antibody (rabbit polyclonal to COX-2, sc-7951) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes
Article Snippet: After treatments, the total protein concentration of cell lysate was determined by BCA (bicinchoninic acid) protein assay kit (Pierce Biotechnology, Rockford, IL) according to manufacture's instruction. .. Membranes were blocked for 1 h with 5% (w/v) nonfat dry milk in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) and were incubated overnight at 4°C in 3% (w/v) BSA with 0.02% (w/v) sodium azide in TBST with p-cPLA2 or cPLA2 antibodies (1:1000 dilution; Cell Signaling Technology, Beverly, MA), anti-IL-1β antibody (1:1000 dilution; R & D Systems, Minneapolis, MN), anti-iNOS antibody (1:1000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin antibody (Sigma, St. Louis, MO).

Transplantation Assay:

Article Title: Endothelial glutathione-S-transferase A4-4 protects against oxidative stress and modulates iNOS expression through NF-?B translocation
Article Snippet: Full-thickness segments of ascending aorta were procured fresh from 7 human donor hearts at the time of cardiac transplantation under an approved human subject protocol (IRB#99-395, University of Texas Medical Branch). .. Lesions and adjacent sections of selected lesions were immunostained for 4-HNE and iNOS with goat anti-HNE antiserum (Alpha Diagnostic, San Antonio, TX), and rabbit anti-iNOS polyclonal antibodies (Santa Cruz Biotechnology., Santa Cruz, CA).

Western Blot:

Article Title: Microglia P2Y6 receptor is related to Parkinson’s disease through neuroinflammatory process
Article Snippet: Paragraph title: Western blotting ... The levels of iNOS and COX-2 were determined using anti-iNOS antibody (sc-69190, 1:500; Santa Cruz) and anti-COX-2 antibodies (160106, 1:800; Caymen Chemical).

Article Title: The Traditional Herbal Medicine, Dangkwisoo-San, Prevents Cerebral Ischemic Injury through Nitric Oxide-Dependent Mechanisms
Article Snippet: Paragraph title: 2.7. Western Blotting ... Immunoblot analysis was performed with anti-eNOS and anti-phospho-eNOS (pS1177) antibodies (BD Biosciences, San Jose, CA), anti-Akt and anti-phospho-Akt (Ser 473) antibodies (Cell signaling, Danvers, MA), anti-nNOS and anti-iNOS antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) followed by incubation with secondary antibody conjugated with horseradish peroxidase.

Article Title: The composition of a bioprocessed shiitake (Lentinus edodes) mushroom mycelia and rice bran formulation and its antimicrobial effects against Salmonella enterica subsp. enterica serovar Typhimurium strain SL1344 in macrophage cells and in mice
Article Snippet: .. Antibodies Western blot analyses were performed using the following antibodies: rabbit anti-mouse iNOS polyclonal antibody from Santa Cruz (Dallas, TX, USA) and mouse anti-Actin monoclonal antibody from Millipore Corp. (Billerica, MA, USA), and all rabbit monoclonal antibodies raised against Beclin-1, Atg5, Atg12, Atg16L, LC3A/B, and phospho-IRF-3 from Cell Signaling Tech. (Danvers, MA, USA). .. Western blot analysis The Western Blot technique was the same as described in previous studies [ , ].

Article Title: The composition of a bioprocessed shiitake (Lentinus edodes) mushroom mycelia and rice bran formulation and its antimicrobial effects against Salmonella enterica subsp. enterica serovar Typhimurium strain SL1344 in macrophage cells and in mice
Article Snippet: .. Western blot analyses were performed using the following antibodies: rabbit anti-mouse iNOS polyclonal antibody from Santa Cruz (Dallas, TX, USA) and mouse anti-Actin monoclonal antibody from Millipore Corp. (Billerica, MA, USA), and all rabbit monoclonal antibodies raised against Beclin-1, Atg5, Atg12, Atg16L, LC3A/B, and phospho-IRF-3 from Cell Signaling Tech. (Danvers, MA, USA). .. The Western Blot technique was the same as described in previous studies [ , ].

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes
Article Snippet: Paragraph title: Western blot analysis ... Membranes were blocked for 1 h with 5% (w/v) nonfat dry milk in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) and were incubated overnight at 4°C in 3% (w/v) BSA with 0.02% (w/v) sodium azide in TBST with p-cPLA2 or cPLA2 antibodies (1:1000 dilution; Cell Signaling Technology, Beverly, MA), anti-IL-1β antibody (1:1000 dilution; R & D Systems, Minneapolis, MN), anti-iNOS antibody (1:1000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin antibody (Sigma, St. Louis, MO).

Article Title: Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles
Article Snippet: Then the cells were harvested for western blotting analysis. .. The separated proteins were then transferred onto polyvinylidene difluoride membrane (Amersham Hybond™ -P, GE Healthcare). iNOS was detected by using monoclonal anti-iNOS antibody NOS2(C-11) at 1:200 (Santa Cruz Biotechnology, Inc.). β-actin was detected using mouse anti- β-actin monoclonal antibody at 1:20,000 (GenScript Corporation).

Immunohistochemistry:

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: .. Immunohistochemical Staining For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v /v ). .. Sections were then washed with PBS and incubated with secondary antibodies.

Article Title: Simvastatin Attenuates the Oxidative Stress, Endothelial Thrombogenicity and the Inducibility of Atrial Fibrillation in a Rat Model of Ischemic Heart Failure
Article Snippet: Paragraph title: 3.7. Immunohistochemistry for eNOS and iNOS ... The sections were incubated with anti-eNOS antibody (Santa Cruz, CA, USA) or anti-iNOS antibody (Santa Cruz, CA, USA) at a 1:50 dilution in PBS.

Article Title: Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome
Article Snippet: Paragraph title: Immunohistochemistry ... Then, sections were incubated with anti-nNOS primer antibody (sc-55521, Santa Cruz Biotechnology, Inc.), anti-iNOS primer antibody (sc-649, Santa Cruz Biotechnology, Inc.), anti-eNOS antibody (sc-654, Santa Cruz Biotechnology, Inc.) in a 1/100 dilution for 18 h at +4°C.

Article Title: Endothelial glutathione-S-transferase A4-4 protects against oxidative stress and modulates iNOS expression through NF-?B translocation
Article Snippet: Paragraph title: Immunohistochemical localization of 4-HNE and iNOS in human atherosclerotic plaques ... Lesions and adjacent sections of selected lesions were immunostained for 4-HNE and iNOS with goat anti-HNE antiserum (Alpha Diagnostic, San Antonio, TX), and rabbit anti-iNOS polyclonal antibodies (Santa Cruz Biotechnology., Santa Cruz, CA).

Article Title: Unique Macrophages Different from M1/M2 Macrophages Inhibit T Cell Mitogenesis while Upregulating Th17 Polarization
Article Snippet: Paragraph title: Histology and immunohistochemistry ... The resultant sections were reacted with rabbit anti-mouse NOS2 Ab (class IgG: Santa Cruz Biotechnology) or rabbit anti-mouse Arg-1 Ab (IgG: Santa Cruz Biotechnology), followed by staining with Alexa Fluor 488- conjugated donkey anti-rabbit IgG Ab (Molecular Probes).

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: .. For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v / v ). .. Sections were then washed with PBS and incubated with secondary antibodies.

Article Title: Significance of vascular endothelial growth factor expression and its correlation with inducible nitric oxide synthase in gastric cancer
Article Snippet: Paragraph title: Immunohistochemical analysis ... The anti-VEGF antibody is a polyclonal mouse anti-serum against VEGF of human origin and recognizes an amino-terminal epitope found in VEGF121,165,189 , and the anti-iNOS antibody is a monoclonal mouse anti-serum against the C-terminal domain of iNOS of human origin (The two antibodies were from Santa Cruz Biotechnology, Inc., CA).

Infection:

Article Title: Unique Macrophages Different from M1/M2 Macrophages Inhibit T Cell Mitogenesis while Upregulating Th17 Polarization
Article Snippet: Histology and immunohistochemistry Spleens excised from mice 2 or 3 weeks after MAC infection were embedded in cryomolds, frozen in liquid nitrogen, cut as serial sections in 14- to 16-μm thickness, and fixed with paraformaldehyde followed by serial treatments with acetic acid/ethanol (1:2) and 1% BSA/0.05% Tween 20 in phosphate-buffered saline (PBS). .. The resultant sections were reacted with rabbit anti-mouse NOS2 Ab (class IgG: Santa Cruz Biotechnology) or rabbit anti-mouse Arg-1 Ab (IgG: Santa Cruz Biotechnology), followed by staining with Alexa Fluor 488- conjugated donkey anti-rabbit IgG Ab (Molecular Probes).

Light Microscopy:

Article Title: Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome
Article Snippet: Then, sections were incubated with anti-nNOS primer antibody (sc-55521, Santa Cruz Biotechnology, Inc.), anti-iNOS primer antibody (sc-649, Santa Cruz Biotechnology, Inc.), anti-eNOS antibody (sc-654, Santa Cruz Biotechnology, Inc.) in a 1/100 dilution for 18 h at +4°C. .. Then, sections were incubated with anti-nNOS primer antibody (sc-55521, Santa Cruz Biotechnology, Inc.), anti-iNOS primer antibody (sc-649, Santa Cruz Biotechnology, Inc.), anti-eNOS antibody (sc-654, Santa Cruz Biotechnology, Inc.) in a 1/100 dilution for 18 h at +4°C.

Article Title: Endothelial glutathione-S-transferase A4-4 protects against oxidative stress and modulates iNOS expression through NF-?B translocation
Article Snippet: Aortic segments were rapidly fixed in 10% neutral-buffered formalin solution and atherosclerotic plaques from 1–4 mm in greatest dimension identified grossly and processed with rare lipid streaks and randomly-sampled adjacent areas of normal-appearing aorta for routine light microscopy. .. Lesions and adjacent sections of selected lesions were immunostained for 4-HNE and iNOS with goat anti-HNE antiserum (Alpha Diagnostic, San Antonio, TX), and rabbit anti-iNOS polyclonal antibodies (Santa Cruz Biotechnology., Santa Cruz, CA).

Inhibition:

Article Title: Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles
Article Snippet: Paragraph title: Inhibition of iNOS expression by cSCKs•siRNA complexes ... The separated proteins were then transferred onto polyvinylidene difluoride membrane (Amersham Hybond™ -P, GE Healthcare). iNOS was detected by using monoclonal anti-iNOS antibody NOS2(C-11) at 1:200 (Santa Cruz Biotechnology, Inc.). β-actin was detected using mouse anti- β-actin monoclonal antibody at 1:20,000 (GenScript Corporation).

Protein Concentration:

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes
Article Snippet: After treatments, the total protein concentration of cell lysate was determined by BCA (bicinchoninic acid) protein assay kit (Pierce Biotechnology, Rockford, IL) according to manufacture's instruction. .. Membranes were blocked for 1 h with 5% (w/v) nonfat dry milk in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) and were incubated overnight at 4°C in 3% (w/v) BSA with 0.02% (w/v) sodium azide in TBST with p-cPLA2 or cPLA2 antibodies (1:1000 dilution; Cell Signaling Technology, Beverly, MA), anti-IL-1β antibody (1:1000 dilution; R & D Systems, Minneapolis, MN), anti-iNOS antibody (1:1000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin antibody (Sigma, St. Louis, MO).

GSH Assay:

Article Title: Green Tea Extract Ameliorates Learning and Memory Deficits in Ischemic Rats via Its Active Component Polyphenol Epigallocatechin-3-gallate by Modulation of Oxidative Stress and Neuroinflammation
Article Snippet: MDA-586 assay Kit and Glutathione (GSH) assay kit were purchased from Cayman Chemical (Ann Arbor, MI, USA). .. Anti-iNOS antibody (rabbit polyclonal to iNOS, sc-651) and anti-COX-2 antibody (rabbit polyclonal to COX-2, sc-7951) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Binding Assay:

Article Title: Simvastatin Attenuates the Oxidative Stress, Endothelial Thrombogenicity and the Inducibility of Atrial Fibrillation in a Rat Model of Ischemic Heart Failure
Article Snippet: Samples were immersed in 10% neutral buffered formalin for 30 min, permeabilized with 0.1% Triton X-100 for 15 min, and antigen retrieval was achieved by heating in 0.01 M citric acid, pH 6.0, for 20 min. Nonspecific binding was blocked with normal goat serum for 15 minutes. .. The sections were incubated with anti-eNOS antibody (Santa Cruz, CA, USA) or anti-iNOS antibody (Santa Cruz, CA, USA) at a 1:50 dilution in PBS.

Immunofluorescence:

Article Title: TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection
Article Snippet: Paragraph title: Histology and immunofluorescence ... Samples were Tissue slides were then incubated overnight at 4°C with a biotin anti-mouse F4/80 antibody (1:150, Abcam) and a polyclonal rabbit anti-mouse iNOS antibody (1:200, Santa Cruz).

Nucleic Acid Electrophoresis:

Article Title: The Traditional Herbal Medicine, Dangkwisoo-San, Prevents Cerebral Ischemic Injury through Nitric Oxide-Dependent Mechanisms
Article Snippet: Proteins were isolated according to standard techniques, separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and transferred onto a nitrocellulose membrane (Amersham Biosciences, Piscataway, NJ). .. Immunoblot analysis was performed with anti-eNOS and anti-phospho-eNOS (pS1177) antibodies (BD Biosciences, San Jose, CA), anti-Akt and anti-phospho-Akt (Ser 473) antibodies (Cell signaling, Danvers, MA), anti-nNOS and anti-iNOS antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) followed by incubation with secondary antibody conjugated with horseradish peroxidase.

Article Title: Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles
Article Snippet: The proteins were subjected to 10% SDS-polyacrylamide gel electrophoresis for detection of β-actin and iNOS, respectively. .. The separated proteins were then transferred onto polyvinylidene difluoride membrane (Amersham Hybond™ -P, GE Healthcare). iNOS was detected by using monoclonal anti-iNOS antibody NOS2(C-11) at 1:200 (Santa Cruz Biotechnology, Inc.). β-actin was detected using mouse anti- β-actin monoclonal antibody at 1:20,000 (GenScript Corporation).

Fluorescence:

Article Title: Unique Macrophages Different from M1/M2 Macrophages Inhibit T Cell Mitogenesis while Upregulating Th17 Polarization
Article Snippet: The resultant sections were reacted with rabbit anti-mouse NOS2 Ab (class IgG: Santa Cruz Biotechnology) or rabbit anti-mouse Arg-1 Ab (IgG: Santa Cruz Biotechnology), followed by staining with Alexa Fluor 488- conjugated donkey anti-rabbit IgG Ab (Molecular Probes). .. Fluorescence microscopy was performed using an FV1000D IX81 confocal microscope (Olympus) and Eclipse 80i fluorescence microscope (Nikon).

Multiple Displacement Amplification:

Article Title: Green Tea Extract Ameliorates Learning and Memory Deficits in Ischemic Rats via Its Active Component Polyphenol Epigallocatechin-3-gallate by Modulation of Oxidative Stress and Neuroinflammation
Article Snippet: MDA-586 assay Kit and Glutathione (GSH) assay kit were purchased from Cayman Chemical (Ann Arbor, MI, USA). .. Anti-iNOS antibody (rabbit polyclonal to iNOS, sc-651) and anti-COX-2 antibody (rabbit polyclonal to COX-2, sc-7951) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Isolation:

Article Title: The Traditional Herbal Medicine, Dangkwisoo-San, Prevents Cerebral Ischemic Injury through Nitric Oxide-Dependent Mechanisms
Article Snippet: Proteins were isolated according to standard techniques, separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and transferred onto a nitrocellulose membrane (Amersham Biosciences, Piscataway, NJ). .. Immunoblot analysis was performed with anti-eNOS and anti-phospho-eNOS (pS1177) antibodies (BD Biosciences, San Jose, CA), anti-Akt and anti-phospho-Akt (Ser 473) antibodies (Cell signaling, Danvers, MA), anti-nNOS and anti-iNOS antibodies (Santa Cruz Biotechnology, Santa Cruz, CA) followed by incubation with secondary antibody conjugated with horseradish peroxidase.

Bicinchoninic Acid Protein Assay:

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes
Article Snippet: After treatments, the total protein concentration of cell lysate was determined by BCA (bicinchoninic acid) protein assay kit (Pierce Biotechnology, Rockford, IL) according to manufacture's instruction. .. Membranes were blocked for 1 h with 5% (w/v) nonfat dry milk in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) and were incubated overnight at 4°C in 3% (w/v) BSA with 0.02% (w/v) sodium azide in TBST with p-cPLA2 or cPLA2 antibodies (1:1000 dilution; Cell Signaling Technology, Beverly, MA), anti-IL-1β antibody (1:1000 dilution; R & D Systems, Minneapolis, MN), anti-iNOS antibody (1:1000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin antibody (Sigma, St. Louis, MO).

Avidin-Biotin Assay:

Article Title: Significance of vascular endothelial growth factor expression and its correlation with inducible nitric oxide synthase in gastric cancer
Article Snippet: The anti-VEGF antibody is a polyclonal mouse anti-serum against VEGF of human origin and recognizes an amino-terminal epitope found in VEGF121,165,189 , and the anti-iNOS antibody is a monoclonal mouse anti-serum against the C-terminal domain of iNOS of human origin (The two antibodies were from Santa Cruz Biotechnology, Inc., CA). .. Location of the primary antibodies was achieved by subsequent application of a biotinylated anti-primary antibody, an avidin - biotin co mplex conju gated to horseradish peroxidase.

Microscopy:

Article Title: Unique Macrophages Different from M1/M2 Macrophages Inhibit T Cell Mitogenesis while Upregulating Th17 Polarization
Article Snippet: The resultant sections were reacted with rabbit anti-mouse NOS2 Ab (class IgG: Santa Cruz Biotechnology) or rabbit anti-mouse Arg-1 Ab (IgG: Santa Cruz Biotechnology), followed by staining with Alexa Fluor 488- conjugated donkey anti-rabbit IgG Ab (Molecular Probes). .. Fluorescence microscopy was performed using an FV1000D IX81 confocal microscope (Olympus) and Eclipse 80i fluorescence microscope (Nikon).

Mouse Assay:

Article Title: Unique Macrophages Different from M1/M2 Macrophages Inhibit T Cell Mitogenesis while Upregulating Th17 Polarization
Article Snippet: Histology and immunohistochemistry Spleens excised from mice 2 or 3 weeks after MAC infection were embedded in cryomolds, frozen in liquid nitrogen, cut as serial sections in 14- to 16-μm thickness, and fixed with paraformaldehyde followed by serial treatments with acetic acid/ethanol (1:2) and 1% BSA/0.05% Tween 20 in phosphate-buffered saline (PBS). .. The resultant sections were reacted with rabbit anti-mouse NOS2 Ab (class IgG: Santa Cruz Biotechnology) or rabbit anti-mouse Arg-1 Ab (IgG: Santa Cruz Biotechnology), followed by staining with Alexa Fluor 488- conjugated donkey anti-rabbit IgG Ab (Molecular Probes).

Lysis:

Article Title: Microglia P2Y6 receptor is related to Parkinson’s disease through neuroinflammatory process
Article Snippet: Western blotting Cells were washed in ice-cold phosphate-buffered saline (PBS) three times and lysed in RIPA lysis buffer (50 mM Tris–HCl, pH 8.0; 1% NP-40; 0.5% sodium deoxycholate; 150 mM NaCl; 0.1% SDS) containing protease and phosphatase inhibitor cocktails (Roche) and 1 mM phenyl-methylsulphonyl fluoride (PMSF). .. The levels of iNOS and COX-2 were determined using anti-iNOS antibody (sc-69190, 1:500; Santa Cruz) and anti-COX-2 antibodies (160106, 1:800; Caymen Chemical).

Article Title: Efficient Protection and Transfection of Small Interfering RNA by Cationic Shell-Crosslinked Knedel-Like Nanoparticles
Article Snippet: The cells were first lysed in 30 μL lysis buffer [1% sodium dodecyl sulfate (SDS), 1.0 mM sodium ortho-vanadate, 10 mM Tris pH 7.4]. .. The separated proteins were then transferred onto polyvinylidene difluoride membrane (Amersham Hybond™ -P, GE Healthcare). iNOS was detected by using monoclonal anti-iNOS antibody NOS2(C-11) at 1:200 (Santa Cruz Biotechnology, Inc.). β-actin was detected using mouse anti- β-actin monoclonal antibody at 1:20,000 (GenScript Corporation).

Software:

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: Immunohistochemical Staining For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v /v ). .. For quantitative analysis, original immunohistochemical photographs were assessed by densitometer using MacBiophotonics ImageJ 1.41a software [ , ].

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v / v ). .. For quantitative analysis, original immunohistochemical photographs were assessed by densitometer using MacBiophotonics ImageJ 1.41a software [ , ].

Article Title: Low energy laser light (632.8 nm) suppresses amyloid-? peptide-induced oxidative and inflammatory responses in astrocytes
Article Snippet: Membranes were blocked for 1 h with 5% (w/v) nonfat dry milk in Tris-buffered saline containing 0.1% (v/v) Tween 20 (TBST) and were incubated overnight at 4°C in 3% (w/v) BSA with 0.02% (w/v) sodium azide in TBST with p-cPLA2 or cPLA2 antibodies (1:1000 dilution; Cell Signaling Technology, Beverly, MA), anti-IL-1β antibody (1:1000 dilution; R & D Systems, Minneapolis, MN), anti-iNOS antibody (1:1000 dilution; Santa Cruz Biotechnology, Santa Cruz, CA) and β-actin antibody (Sigma, St. Louis, MO). .. The protein bands detected on x-ray film were quantified using a computer-driven scanner and Quantity One software (Bio-Rad, Hercules, CA).

Laser-Scanning Microscopy:

Article Title: TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection
Article Snippet: Samples were Tissue slides were then incubated overnight at 4°C with a biotin anti-mouse F4/80 antibody (1:150, Abcam) and a polyclonal rabbit anti-mouse iNOS antibody (1:200, Santa Cruz). .. Samples were analyzed using the Zeiss 510 LSM Meta confocal laser scanning microscope and Zeiss LSM Image Browser program (Carl Zeiss, Hudson, OH, USA).

Formalin-fixed Paraffin-Embedded:

Article Title: Simvastatin Attenuates the Oxidative Stress, Endothelial Thrombogenicity and the Inducibility of Atrial Fibrillation in a Rat Model of Ischemic Heart Failure
Article Snippet: Briefly, formalin-fixed, paraffin-embedded tissue was prepared using conventional histological methods. .. The sections were incubated with anti-eNOS antibody (Santa Cruz, CA, USA) or anti-iNOS antibody (Santa Cruz, CA, USA) at a 1:50 dilution in PBS.

Article Title: Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome
Article Snippet: Immunohistochemistry Formalin-fixed, paraffin-embedded sections were used for immunohistochemical staining. .. Then, sections were incubated with anti-nNOS primer antibody (sc-55521, Santa Cruz Biotechnology, Inc.), anti-iNOS primer antibody (sc-649, Santa Cruz Biotechnology, Inc.), anti-eNOS antibody (sc-654, Santa Cruz Biotechnology, Inc.) in a 1/100 dilution for 18 h at +4°C.

Staining:

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: .. Immunohistochemical Staining For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v /v ). .. Sections were then washed with PBS and incubated with secondary antibodies.

Article Title: Effect of nitrergic system on colonic motility in a rat model of irritable bowel syndrome
Article Snippet: Immunohistochemistry Formalin-fixed, paraffin-embedded sections were used for immunohistochemical staining. .. Then, sections were incubated with anti-nNOS primer antibody (sc-55521, Santa Cruz Biotechnology, Inc.), anti-iNOS primer antibody (sc-649, Santa Cruz Biotechnology, Inc.), anti-eNOS antibody (sc-654, Santa Cruz Biotechnology, Inc.) in a 1/100 dilution for 18 h at +4°C.

Article Title: Endothelial glutathione-S-transferase A4-4 protects against oxidative stress and modulates iNOS expression through NF-?B translocation
Article Snippet: Sections (4 μm) were stained with hematoxylin and eosin (H+E) and atherosclerotic plaques graded according to established criteria. ( ) From 2 to 6 plaques were identified in each patient’s aortic sample, giving an approximate total of 32 plaques of varying ages examined. .. Lesions and adjacent sections of selected lesions were immunostained for 4-HNE and iNOS with goat anti-HNE antiserum (Alpha Diagnostic, San Antonio, TX), and rabbit anti-iNOS polyclonal antibodies (Santa Cruz Biotechnology., Santa Cruz, CA).

Article Title: TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection
Article Snippet: Histology and immunofluorescence A longitudinal strip from the greater curvature of the stomach was excised and placed in 10% normal buffered formalin for 24 h, embedded in paraffin and processed routinely for hematoxylin and eosin (H & E) or immunofluorescence staining. .. Samples were Tissue slides were then incubated overnight at 4°C with a biotin anti-mouse F4/80 antibody (1:150, Abcam) and a polyclonal rabbit anti-mouse iNOS antibody (1:200, Santa Cruz).

Article Title: Unique Macrophages Different from M1/M2 Macrophages Inhibit T Cell Mitogenesis while Upregulating Th17 Polarization
Article Snippet: .. The resultant sections were reacted with rabbit anti-mouse NOS2 Ab (class IgG: Santa Cruz Biotechnology) or rabbit anti-mouse Arg-1 Ab (IgG: Santa Cruz Biotechnology), followed by staining with Alexa Fluor 488- conjugated donkey anti-rabbit IgG Ab (Molecular Probes). .. Fluorescence microscopy was performed using an FV1000D IX81 confocal microscope (Olympus) and Eclipse 80i fluorescence microscope (Nikon).

Article Title: Neuroprotective Effects of dl-3-n-Butylphthalide against Doxorubicin-Induced Neuroinflammation, Oxidative Stress, Endoplasmic Reticulum Stress, and Behavioral Changes
Article Snippet: Paragraph title: 2.9. Immunohistochemical Staining ... For immunohistochemical, hippocampus sections were incubated overnight with anti-iNOS antibody (Santa Cruz Biotechnology, 1 : 500 in PBS, v / v ).

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  • 93
    Santa Cruz Biotechnology anti inos
    Histological observation of rat hind footpads after injecting Carr 0.9% saline (control group) or Carr. (a-d) H E staining of footpad tissue sections from rat in each group. (e-h) <t>iNOS</t> immunohistochemical staining of footpad tissue sections from rat in each group. (i-l) <t>COX-2</t> immunohistochemical staining of footpad tissue sections from rat in each group. Scale bar = 50 µ m. The infiltrating cells were predominantly neutrophils (N; arrows). The brown staining indicates the interaction of primary and secondary antibodies and the presence of iNOS and COX-2.
    Anti Inos, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Histological observation of rat hind footpads after injecting Carr 0.9% saline (control group) or Carr. (a-d) H E staining of footpad tissue sections from rat in each group. (e-h) iNOS immunohistochemical staining of footpad tissue sections from rat in each group. (i-l) COX-2 immunohistochemical staining of footpad tissue sections from rat in each group. Scale bar = 50 µ m. The infiltrating cells were predominantly neutrophils (N; arrows). The brown staining indicates the interaction of primary and secondary antibodies and the presence of iNOS and COX-2.

    Journal: BioMed Research International

    Article Title: Anti-Inflammatory Activities of Leaf Oil from Cinnamomum subavenium In Vitro and In Vivo

    doi: 10.1155/2019/1823149

    Figure Lengend Snippet: Histological observation of rat hind footpads after injecting Carr 0.9% saline (control group) or Carr. (a-d) H E staining of footpad tissue sections from rat in each group. (e-h) iNOS immunohistochemical staining of footpad tissue sections from rat in each group. (i-l) COX-2 immunohistochemical staining of footpad tissue sections from rat in each group. Scale bar = 50 µ m. The infiltrating cells were predominantly neutrophils (N; arrows). The brown staining indicates the interaction of primary and secondary antibodies and the presence of iNOS and COX-2.

    Article Snippet: Anti-iNOS, anti-COX-2 antibody, and anti-NF-κ B p65 antibody were from Santa Cruz Biotechnology (USA).

    Techniques: Staining, Immunohistochemistry

    The effects of steppogenin ( 1 ) on nitrite ( A ) production and iNOS and COX-2 expression ( B ) in lipopolysaccharide (LPS)-stimulated primary rat microglial cells. ( A , B ) The cells were pretreated for 3 h with the indicated concentrations of 1 and then stimulated for 24 h with LPS (1 μg/mL). The data are presented as the mean ± SD of three experiments. The band intensities were quantified by densitometry and normalized to the intensities of the β-actin band; the normalized values are presented below each band. ** p

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: Steppogenin Isolated from Cudrania tricuspidata Shows Antineuroinflammatory Effects via NF-κB and MAPK Pathways in LPS-Stimulated BV2 and Primary Rat Microglial Cells

    doi: 10.3390/molecules22122130

    Figure Lengend Snippet: The effects of steppogenin ( 1 ) on nitrite ( A ) production and iNOS and COX-2 expression ( B ) in lipopolysaccharide (LPS)-stimulated primary rat microglial cells. ( A , B ) The cells were pretreated for 3 h with the indicated concentrations of 1 and then stimulated for 24 h with LPS (1 μg/mL). The data are presented as the mean ± SD of three experiments. The band intensities were quantified by densitometry and normalized to the intensities of the β-actin band; the normalized values are presented below each band. ** p

    Article Snippet: Primary antibodies, including mouse/goat/rabbit anti-COX-2 (sc-1745), anti-iNOS (sc-650), anti-β-actin (sc-47778), anti-IкB-α (sc-371), anti-phospho-IкB-α (sc-8404), anti-p50 (sc-7178), anti-p65 (sc-8008), and anti-proliferating cell nuclear antigen (PCNA) (sc-7907), and secondary antibodies were purchased from Santa Cruz Biotechnology (Heidelberg, Germany).

    Techniques: Expressing

    The effects of steppogenin ( 1 ) on nitrite ( A ) and prostaglandin E2 (PGE 2 ) ( B ) production and iNOS and COX-2 expression ( C ) in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. ( A – C ) The cells were pretreated for 3 h with the indicated concentrations of 1 and then stimulated for 24 h with LPS (1 μg/mL). The data are presented as the mean ± SD of three experiments. The band intensity was quantified by densitometry and normalized to the intensity of the β-actin band; the normalized values are presented below each band. * p

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: Steppogenin Isolated from Cudrania tricuspidata Shows Antineuroinflammatory Effects via NF-κB and MAPK Pathways in LPS-Stimulated BV2 and Primary Rat Microglial Cells

    doi: 10.3390/molecules22122130

    Figure Lengend Snippet: The effects of steppogenin ( 1 ) on nitrite ( A ) and prostaglandin E2 (PGE 2 ) ( B ) production and iNOS and COX-2 expression ( C ) in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. ( A – C ) The cells were pretreated for 3 h with the indicated concentrations of 1 and then stimulated for 24 h with LPS (1 μg/mL). The data are presented as the mean ± SD of three experiments. The band intensity was quantified by densitometry and normalized to the intensity of the β-actin band; the normalized values are presented below each band. * p

    Article Snippet: Primary antibodies, including mouse/goat/rabbit anti-COX-2 (sc-1745), anti-iNOS (sc-650), anti-β-actin (sc-47778), anti-IкB-α (sc-371), anti-phospho-IкB-α (sc-8404), anti-p50 (sc-7178), anti-p65 (sc-8008), and anti-proliferating cell nuclear antigen (PCNA) (sc-7907), and secondary antibodies were purchased from Santa Cruz Biotechnology (Heidelberg, Germany).

    Techniques: Expressing

    Inhibition of iNOS and COX-2 protein expression by scopoletin induced by Carr in mice paw edema for 5th hour. Normal control received 0.9% normal saline. Animals treated with scopoletin (1, 5, and 10 mg/kg) and Indo to injection of Carr right hind paws. The right hind paw tissues were taken at the 5 hour. Then the homogenate was centrifuged and tissue suspended were then prepared and subjected to western blotting using an antibody specific for iNOS and COX-2. β -actin was used as an internal control. (a) Representative western blot from two separate experiments was shown. (b) Relative iNOS and COX-2 protein levels were calculated with reference to Carr-injected mouse. Each point represents the average value for three individual animals . ### compared with sample of control group. The data were presented as mean ± S.E.M. for three different experiments performed in triplicate. ** P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Ameliorative Effects of Scopoletin from Crossostephium chinensis against Inflammation Pain and Its Mechanisms in Mice

    doi: 10.1155/2012/595603

    Figure Lengend Snippet: Inhibition of iNOS and COX-2 protein expression by scopoletin induced by Carr in mice paw edema for 5th hour. Normal control received 0.9% normal saline. Animals treated with scopoletin (1, 5, and 10 mg/kg) and Indo to injection of Carr right hind paws. The right hind paw tissues were taken at the 5 hour. Then the homogenate was centrifuged and tissue suspended were then prepared and subjected to western blotting using an antibody specific for iNOS and COX-2. β -actin was used as an internal control. (a) Representative western blot from two separate experiments was shown. (b) Relative iNOS and COX-2 protein levels were calculated with reference to Carr-injected mouse. Each point represents the average value for three individual animals . ### compared with sample of control group. The data were presented as mean ± S.E.M. for three different experiments performed in triplicate. ** P

    Article Snippet: Anti-iNOS, anti-COX-2, and anti-β -actin antibody (Santa Cruz, USA), and a protein assay kit (Bio-Rad Laboratories Ltd., Watford, Hertfordshire, UK) were obtained as indicated.

    Techniques: Inhibition, Expressing, Mouse Assay, Injection, Western Blot

    CAMK4 is necessary for crocin-mediated inhibition of iNOS expression in LPS-stimulated macrophages. RAW 264.7 cells were transfected with CAMK4 siRNA or control siRNA and then treated with crocin (500 μ M). After 3 h, cells were incubated with LPS (0.1 μ g/mL) for 24 h. Equal amounts of cytosolic extract were analyzed by Western blotting. Tubulin was used as a loading control.

    Journal: Mediators of Inflammation

    Article Title: Crocin Suppresses LPS-Stimulated Expression of Inducible Nitric Oxide Synthase by Upregulation of Heme Oxygenase-1 via Calcium/Calmodulin-Dependent Protein Kinase 4

    doi: 10.1155/2014/728709

    Figure Lengend Snippet: CAMK4 is necessary for crocin-mediated inhibition of iNOS expression in LPS-stimulated macrophages. RAW 264.7 cells were transfected with CAMK4 siRNA or control siRNA and then treated with crocin (500 μ M). After 3 h, cells were incubated with LPS (0.1 μ g/mL) for 24 h. Equal amounts of cytosolic extract were analyzed by Western blotting. Tubulin was used as a loading control.

    Article Snippet: The gel was then transferred to 0.45 μ m nitrocellulose paper and incubated with anti-iNOS, p65, HO-1, Nrf2, phospho-CAMK4, Akt, ERK1/2, JNK, TBP, HDAC antibodies (Santa Cruz Biotechnology, Santa Cruz, CA, USA), CAMK4, phospho-Akt, phospho-ERK1/2, phospho-JNK antibodies (Cell Signaling Technology, Beverly, MA, USA) or α -tubulin antibody (Bio Genex, Fremont, CA, USA), and secondary antibody and then detected by an enhanced chemiluminescence detection system according to the recommended procedure (GE Healthcare, Piscataway, NJ, USA).

    Techniques: Inhibition, Expressing, Transfection, Incubation, Western Blot