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  • 99
    Thermo Fisher tumor necrosis factor α tnf α
    Attenuation of the level of reactive oxygen species (ROS) by genistein. (A) Keratinocytes were stimulated respectively with a proinflammatory “cytokine mix” corresponding to a concentration of 2 ng/mL (ACT 2) or 5 ng/mL (ACT 5) in each compound of the mix and incubated with 10 mM N-acetyl-cysteine (NAC) (ACT 2 + NAC), 100 μM genistein (ACT 2 + GEN), 10 ng/mL <t>TNF-α</t> or 1 μg/mL LPS alone, and with 10 ng/mL TNF-α or 1 μg/mL LPS incubated with 100 μM genistein (TNF-α + GEN or LPS + GEN). DMSO-treated, unstimulated cells were used as the control (NACT). Additional control was used in the form of unstimulated cells treated with 100 μM genistein (GEN). Intracellular ROS levels were examined by a CellROX Deep Red Reagent and confocal fluorescence microscopy. Nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments (with scale bars 25 μm) are shown. (B) Analysis of ROS were additionally performed by fluorescent cell analyzer. The data are presented as the means ± standard deviation (SD) from three independent experiments. Significant differences ( p ≤ 0.05) between cell populations not expressing intracellular ROS (ROS [–]) and expressing intracellular ROS (ROS [+]) were observed for all tested conditions, except for LPS where cell numbers of ROS (-) and ROS (+) were comparable. Most important, statistically significant differences of p ≤ 0.05 within ROS groups are indicated with * for samples of TNF-α, TNF-α + GEN, and LPS versus NACT, with † for sample TNF-α + GEN with respect to TNF-α, while with ‡ for sample LPS + GEN referred to LPS. Statistical analysis was performed using ANOVA with Tukey’s HSD test.
    Tumor Necrosis Factor α Tnf α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 945 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
    Tumor Necrosis Factor α Tnf α, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 538 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    R&D Systems tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
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    93
    Becton Dickinson tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
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    93
    Santa Cruz Biotechnology tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
    Tumor Necrosis Factor α Tnf α, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 182 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    BioLegend tumour necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
    Tumour Necrosis Factor α Tnf α, supplied by BioLegend, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ApexBio tumour necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
    Tumour Necrosis Factor α Tnf α, supplied by ApexBio, 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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    92
    USCN Life tumor necrosis factor α tnf α
    TM5441 pre-treatment and siPAI-1 improved <t>TNF-α-induced</t> mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p
    Tumor Necrosis Factor α Tnf α, supplied by USCN Life, used in various techniques. Bioz Stars score: 92/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tnf α, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1810 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Haematologic Technologies tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
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    93
    Bio-Rad tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 255 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Roche tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cusabio tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
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    Boehringer Mannheim tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    GENTAUR tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by GENTAUR, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Proteintech tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by Proteintech, used in various techniques. Bioz Stars score: 94/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    VECTOR-BEST tumour necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumour Necrosis Factor α Tnf α, supplied by VECTOR-BEST, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PeproTech tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
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    Elabscience tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
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    GE Healthcare tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cloud-Clone tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by Cloud-Clone, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boster Bio tumor necrosis factor α tnf α
    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and <t>TNF-</t> α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).
    Tumor Necrosis Factor α Tnf α, supplied by Boster Bio, used in various techniques. Bioz Stars score: 93/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Attenuation of the level of reactive oxygen species (ROS) by genistein. (A) Keratinocytes were stimulated respectively with a proinflammatory “cytokine mix” corresponding to a concentration of 2 ng/mL (ACT 2) or 5 ng/mL (ACT 5) in each compound of the mix and incubated with 10 mM N-acetyl-cysteine (NAC) (ACT 2 + NAC), 100 μM genistein (ACT 2 + GEN), 10 ng/mL TNF-α or 1 μg/mL LPS alone, and with 10 ng/mL TNF-α or 1 μg/mL LPS incubated with 100 μM genistein (TNF-α + GEN or LPS + GEN). DMSO-treated, unstimulated cells were used as the control (NACT). Additional control was used in the form of unstimulated cells treated with 100 μM genistein (GEN). Intracellular ROS levels were examined by a CellROX Deep Red Reagent and confocal fluorescence microscopy. Nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments (with scale bars 25 μm) are shown. (B) Analysis of ROS were additionally performed by fluorescent cell analyzer. The data are presented as the means ± standard deviation (SD) from three independent experiments. Significant differences ( p ≤ 0.05) between cell populations not expressing intracellular ROS (ROS [–]) and expressing intracellular ROS (ROS [+]) were observed for all tested conditions, except for LPS where cell numbers of ROS (-) and ROS (+) were comparable. Most important, statistically significant differences of p ≤ 0.05 within ROS groups are indicated with * for samples of TNF-α, TNF-α + GEN, and LPS versus NACT, with † for sample TNF-α + GEN with respect to TNF-α, while with ‡ for sample LPS + GEN referred to LPS. Statistical analysis was performed using ANOVA with Tukey’s HSD test.

    Journal: PLoS ONE

    Article Title: Molecular action of isoflavone genistein in the human epithelial cell line HaCaT

    doi: 10.1371/journal.pone.0192297

    Figure Lengend Snippet: Attenuation of the level of reactive oxygen species (ROS) by genistein. (A) Keratinocytes were stimulated respectively with a proinflammatory “cytokine mix” corresponding to a concentration of 2 ng/mL (ACT 2) or 5 ng/mL (ACT 5) in each compound of the mix and incubated with 10 mM N-acetyl-cysteine (NAC) (ACT 2 + NAC), 100 μM genistein (ACT 2 + GEN), 10 ng/mL TNF-α or 1 μg/mL LPS alone, and with 10 ng/mL TNF-α or 1 μg/mL LPS incubated with 100 μM genistein (TNF-α + GEN or LPS + GEN). DMSO-treated, unstimulated cells were used as the control (NACT). Additional control was used in the form of unstimulated cells treated with 100 μM genistein (GEN). Intracellular ROS levels were examined by a CellROX Deep Red Reagent and confocal fluorescence microscopy. Nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Results representative of three independent experiments (with scale bars 25 μm) are shown. (B) Analysis of ROS were additionally performed by fluorescent cell analyzer. The data are presented as the means ± standard deviation (SD) from three independent experiments. Significant differences ( p ≤ 0.05) between cell populations not expressing intracellular ROS (ROS [–]) and expressing intracellular ROS (ROS [+]) were observed for all tested conditions, except for LPS where cell numbers of ROS (-) and ROS (+) were comparable. Most important, statistically significant differences of p ≤ 0.05 within ROS groups are indicated with * for samples of TNF-α, TNF-α + GEN, and LPS versus NACT, with † for sample TNF-α + GEN with respect to TNF-α, while with ‡ for sample LPS + GEN referred to LPS. Statistical analysis was performed using ANOVA with Tukey’s HSD test.

    Article Snippet: For further experiments, cells were stimulated with a combination of a proinflammatory “cytokine mix”: IL-1A, IL-17A, IL-22, oncostatin M (OSM), and tumor necrosis factor-α (TNF-α) (Gibco, Thermo Fisher Scientific, CA, USA) 2 ng/mL each or lipopolysaccharide (LPS, from Escherichia coli 055 :B5 , Sigma-Aldrich, St. Louis, USA) 1 μg/mL in the presence or absence of genistein for 24 hours [ ].

    Techniques: Concentration Assay, Activated Clotting Time Assay, Incubation, Fluorescence, Microscopy, Staining, Standard Deviation, Expressing

    TM5441 pre-treatment and siPAI-1 improved TNF-α-induced mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p

    Journal: Oncotarget

    Article Title: TM5441, a plasminogen activator inhibitor-1 inhibitor, protects against high fat diet-induced non-alcoholic fatty liver disease

    doi: 10.18632/oncotarget.21120

    Figure Lengend Snippet: TM5441 pre-treatment and siPAI-1 improved TNF-α-induced mitochondrial dysfunction in HepG2 cells HepG2 cells were pretreated with 20 μM TM5441 before the exposure of 50 ng/ml TNF-α for 24 hours. mRNA expressions of ( A ) PAI-1 and ( B) mitochondria biogenesis-related genes, such as PGC-1α, mtDNA, TFAM, NRF1, and NRF2 were measured by RT-qPCR. Then, PAI-1 is downregulated with siRNA in which confirmed by immunoblot analysis. ( C ) 20 nM siPAI-1 was transfected 24 hours before the exposure of 50 ng/ml TNF-α for 24 hour. The mRNA gene expression of ( D ) PAI-1 and ( E ) the mitochondrial function-related genes were measured by RT-qPCR. Data are presented as mean±SE of 4-5 experiments. * p

    Article Snippet: Growth arrested and synchronized cells were exposed with 50 ng/mL tumor necrosis factor-α/TNF-α (Sigma-Aldrich, St. Louis, MO, USA) or 25 and 50 nM mouse recombinant PAI-1 (EMD Millipore, Billerica, MA, USA) in the presence and absence of 20 μM TM5441.

    Techniques: Pyrolysis Gas Chromatography, Quantitative RT-PCR, Transfection, Expressing

    Early TM5441 treatment suppressed hepatic inflammation in HFD mice ( A , B ) Anti-inflammatory effect of TM5441 was analyzed through quantifying positive F4/80 IHC staining area in the liver section. Magnification, 200×; scale bar, 50 μm. Pro-inflammatory mRNA expressions in the liver, such as ( C ) F4/80, ( D ) MCP-1 ( E ) TNF-α, ( F ) NLRP3, were measured by RT-qPCR. Data are shown as mean ± SE of 7–8 mice. * p

    Journal: Oncotarget

    Article Title: TM5441, a plasminogen activator inhibitor-1 inhibitor, protects against high fat diet-induced non-alcoholic fatty liver disease

    doi: 10.18632/oncotarget.21120

    Figure Lengend Snippet: Early TM5441 treatment suppressed hepatic inflammation in HFD mice ( A , B ) Anti-inflammatory effect of TM5441 was analyzed through quantifying positive F4/80 IHC staining area in the liver section. Magnification, 200×; scale bar, 50 μm. Pro-inflammatory mRNA expressions in the liver, such as ( C ) F4/80, ( D ) MCP-1 ( E ) TNF-α, ( F ) NLRP3, were measured by RT-qPCR. Data are shown as mean ± SE of 7–8 mice. * p

    Article Snippet: Growth arrested and synchronized cells were exposed with 50 ng/mL tumor necrosis factor-α/TNF-α (Sigma-Aldrich, St. Louis, MO, USA) or 25 and 50 nM mouse recombinant PAI-1 (EMD Millipore, Billerica, MA, USA) in the presence and absence of 20 μM TM5441.

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Quantitative RT-PCR

    Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and TNF- α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).

    Journal: Experimental cell research

    Article Title: Characterization of the anti-angiogenic properties of arresten, an ?1?1 integrin dependent collagen-derived tumor suppressor

    doi: 10.1016/j.yexcr.2008.08.011

    Figure Lengend Snippet: Arresten does not induce apoptosis in carcinoma, fibrosarcoma or fibroblast cell lines in vitro Arresten did not increase the activity of caspase-3 in HSC-3 oral squamous cell carcinoma cells ( A ), PC-3 prostate adenocarcinoma cells ( B ), 786-0 renal cell carcinoma cells ( C ), HT1080 fibrosarcoma cells ( D ) or primary gingival fibroblasts ( E ). Control cells received only PBS, and TNF- α (40 ng/ml) was used as a positive control for apoptosis. DEVD-fmk, a specific caspase-3 inhibitor (inhib), was used to ensure the specificity of the assay. The caspase-3 assay was performed twice ( A, D, E ) or once ( B–C ).

    Article Snippet: After a 24-h incubation with arresten (700 nM) or TNF-α (40 ng/ml) in low-serum media (2%), the HMVEC endothelial cells (90% confluent) were washed with cold PBS, lysed on ice in 1× lysis buffer (Cell Signaling) for 5 min, briefly sonicated and centrifuged for 10 min at 14000×g at 4°C.

    Techniques: In Vitro, Activity Assay, Positive Control, Inhibition, Caspase-3 Assay

    Increased apoptosis after arresten treatment in vitro and in vivo in the TUNEL assay A-E. 0.5 × 10 6 C-PAE cells in 6-well plates were treated with arresten (350 and 700 nM), TNF- α (40 ng/ml) or PBS in media containing 3 ng/ml bFGF and 5 ng/ml VEGF. The cells were then collected and the TUNEL assay was performed. Apoptotic cells are shown in green (TdT(+) staining, A–B ) and all cells are stained with red (propidium iodine, C–D ). Arresten treatment ( B ) significantly increases the number of apoptotic cells, when compared to PBS treatment ( A ). E. The number of apoptotic cells per high power field was counted. Both arresten and TNF- α treatments significantly increase the number of apoptotic cells, when compared to cells treated with PBS (*** p

    Journal: Experimental cell research

    Article Title: Characterization of the anti-angiogenic properties of arresten, an ?1?1 integrin dependent collagen-derived tumor suppressor

    doi: 10.1016/j.yexcr.2008.08.011

    Figure Lengend Snippet: Increased apoptosis after arresten treatment in vitro and in vivo in the TUNEL assay A-E. 0.5 × 10 6 C-PAE cells in 6-well plates were treated with arresten (350 and 700 nM), TNF- α (40 ng/ml) or PBS in media containing 3 ng/ml bFGF and 5 ng/ml VEGF. The cells were then collected and the TUNEL assay was performed. Apoptotic cells are shown in green (TdT(+) staining, A–B ) and all cells are stained with red (propidium iodine, C–D ). Arresten treatment ( B ) significantly increases the number of apoptotic cells, when compared to PBS treatment ( A ). E. The number of apoptotic cells per high power field was counted. Both arresten and TNF- α treatments significantly increase the number of apoptotic cells, when compared to cells treated with PBS (*** p

    Article Snippet: After a 24-h incubation with arresten (700 nM) or TNF-α (40 ng/ml) in low-serum media (2%), the HMVEC endothelial cells (90% confluent) were washed with cold PBS, lysed on ice in 1× lysis buffer (Cell Signaling) for 5 min, briefly sonicated and centrifuged for 10 min at 14000×g at 4°C.

    Techniques: In Vitro, In Vivo, TUNEL Assay, Staining

    Arresten induces endothelial cell apoptosis Annexin V-FITC staining was performed on C-PAE treated with 175 nM arresten for 0, 2 or 4 hours. Control cells received only PBS, and TNF- α (40 ng/ml) was used as a positive control for apoptosis. A–B. Arresten treatment induced morphological changes typical for apoptosis in the C-PAE cells ( B ) when compared to the control cells treated with PBS ( A ). C. The percentage of annexin V positive cells was plotted after different time points (0, 2 and 4 hours). Arresten induced apoptosis at a similar rate to TNF- α. This induction of apoptosis is statistically significant when compared to the cells treated with PBS (* p

    Journal: Experimental cell research

    Article Title: Characterization of the anti-angiogenic properties of arresten, an ?1?1 integrin dependent collagen-derived tumor suppressor

    doi: 10.1016/j.yexcr.2008.08.011

    Figure Lengend Snippet: Arresten induces endothelial cell apoptosis Annexin V-FITC staining was performed on C-PAE treated with 175 nM arresten for 0, 2 or 4 hours. Control cells received only PBS, and TNF- α (40 ng/ml) was used as a positive control for apoptosis. A–B. Arresten treatment induced morphological changes typical for apoptosis in the C-PAE cells ( B ) when compared to the control cells treated with PBS ( A ). C. The percentage of annexin V positive cells was plotted after different time points (0, 2 and 4 hours). Arresten induced apoptosis at a similar rate to TNF- α. This induction of apoptosis is statistically significant when compared to the cells treated with PBS (* p

    Article Snippet: After a 24-h incubation with arresten (700 nM) or TNF-α (40 ng/ml) in low-serum media (2%), the HMVEC endothelial cells (90% confluent) were washed with cold PBS, lysed on ice in 1× lysis buffer (Cell Signaling) for 5 min, briefly sonicated and centrifuged for 10 min at 14000×g at 4°C.

    Techniques: Staining, Positive Control