mouse anti human par2 antibody  (Santa Cruz Biotechnology)

Journal: Cellular and Molecular Neurobiology

Article Title: Procoagulant Extracellular Vesicles Increase Neuronal Tau expression, Metabolism and Processing Through Tissue Factor and Protease Activated Receptor 2

doi: 10.1007/s10571-025-01658-7

Figure Lengend Snippet: Examination of the mechanism of influence of TF on Tau protein expression and phosphorylation in differentiated SH-SY5Y cells. SH-SY5Y (2 × 10 5 ) were treated with recombinant relipidated Innovin TF (0.65 ng/ml) together with or without human fVIIa (5 nM) or fVIIa alone. In some experiments, the TF aliquots were pre-incubated for 1 h, with 10H10 antibody (20 µg/ml) capable of blocking TF signalling, HTF-1 antibody (20 µg/ml) to block TF-fVIIa protease/procoagulant activity, or a mouse control isotype IgG antibody (20 µg/ml; not shown). In other experiments, fVIIa was pre-incubated for 1 h with the chemical inhibitor PCI27483 (10 µg/ml). Alternatively, the neuronal cells were treated with AIIB2 antibody (20 µg/ml) to block β1-integrin signalling, SAM11 antibody (20 µg/ml) capable of blocking PAR2 signalling, or PAR2-activating peptide (PAR2-AP; 20 µM) to induce PAR2 signalling. Cells were harvested at 24 h, and cellular lysates (10 µg protein) were examined for Tau and phospho-Thr181 Tau by western blot analysis. All values were normalised against the respective GAPDH and for comparison, all ratios were calculated against the average from the non-treated cells ± the calculated standard deviation. The number of experiments is shown in each column, and all data groups were determined to have normal distributions and are shown in each column. A SH-SY5Y protein electrophoresis, B SH-SY5Y phospho-Thr181 Tau electrophoresis, C SH-SY5Y Tau protein ratio, and D SH-SY5Y phospho-Thr181 Tau ratio

Article Snippet: Alternatively, the neuronal cells were treated with a rat anti-human antibody (20 μg/ml; AIIB2; Merck KGaA) to block β1-integrin signalling, a mouse anti-human PAR2 antibody, SAM11 (20 μg/ml; Santa Cruz Biotechnology, Heidelberg, Germany), capable of blocking PAR2 signalling, or PAR2-activating peptide (PAR2-AP; 20 μM) to induce PAR2 signalling.

Techniques: Expressing, Phospho-proteomics, Recombinant, Incubation, Blocking Assay, Activity Assay, Control, Western Blot, Comparison, Standard Deviation, Protein Electrophoresis, Electrophoresis

Journal: Cellular and Molecular Neurobiology

Article Title: Procoagulant Extracellular Vesicles Increase Neuronal Tau expression, Metabolism and Processing Through Tissue Factor and Protease Activated Receptor 2

doi: 10.1007/s10571-025-01658-7

Figure Lengend Snippet: Time-course analysis of the Tau protein fragments in differentiated SH-SY5Y cells, following treatment with TF . SH-SY5Y (2 × 10 5 ) were treated with as single dose of recombinant relipidated Innovin TF (0.65 ng/ml) together with human fVIIa (5 nM). In some experiments, the TF aliquots were pre-incubated for 1 h, with 10H10 antibody (20 µg/ml) capable of blocking TF signalling, HTF-1 antibody (20 µg/ml) to block TF-fVIIa protease/procoagulant activity, or a mouse control isotype IgG antibody (20 µg/ml; not shown). Alternatively, the neuronal cells were treated with AIIB2 antibody (20 µg/ml) to block β1-integrin signalling, or SAM11 antibody (20 µg/ml) capable of blocking PAR2 signalling. Sets of cells were harvested at A 48 h and D at 72 h and cellular lysates (10 µg protein) were examined for Tau by western blot analysis. All values were normalised against the respective GAPDH (see Supplementary Fig. 6 A and B) and for comparison, all ratios were calculated against the average from the non-treated cells ± the calculated standard deviation. The data were obtained from 6 biological experiments, and all data groups were determined to have normal distributions. A Electrophoresis at 48 h, B calculated ratios of 50 kDa bands at 48 h, C calculated ratios of 30–35 kDa bands at 48 h, D electrophoresis at 72 h, E calculated ratios of 50 kDa bands at 72 h, F calculated ratios of 40 kDa bands at 72 h, G calculated ratios of 30–35 kDa bands at 72 h

Article Snippet: Alternatively, the neuronal cells were treated with a rat anti-human antibody (20 μg/ml; AIIB2; Merck KGaA) to block β1-integrin signalling, a mouse anti-human PAR2 antibody, SAM11 (20 μg/ml; Santa Cruz Biotechnology, Heidelberg, Germany), capable of blocking PAR2 signalling, or PAR2-activating peptide (PAR2-AP; 20 μM) to induce PAR2 signalling.

Techniques: Recombinant, Incubation, Blocking Assay, Activity Assay, Control, Western Blot, Comparison, Standard Deviation, Electrophoresis

Journal: Cellular and Molecular Neurobiology

Article Title: Procoagulant Extracellular Vesicles Increase Neuronal Tau expression, Metabolism and Processing Through Tissue Factor and Protease Activated Receptor 2

doi: 10.1007/s10571-025-01658-7

Figure Lengend Snippet: Examination of the mechanism of influence of TF on Tau protein expression and phosphorylation in differentiated SH-SY5Y cells. SH-SY5Y (2 × 10 5 ) were treated with recombinant relipidated Innovin TF (0.65 ng/ml) together with or without human fVIIa (5 nM) or fVIIa alone. In some experiments, the TF aliquots were pre-incubated for 1 h, with 10H10 antibody (20 µg/ml) capable of blocking TF signalling, HTF-1 antibody (20 µg/ml) to block TF-fVIIa protease/procoagulant activity, or a mouse control isotype IgG antibody (20 µg/ml; not shown). In other experiments, fVIIa was pre-incubated for 1 h with the chemical inhibitor PCI27483 (10 µg/ml). Alternatively, the neuronal cells were treated with AIIB2 antibody (20 µg/ml) to block β1-integrin signalling, SAM11 antibody (20 µg/ml) capable of blocking PAR2 signalling, or PAR2-activating peptide (PAR2-AP; 20 µM) to induce PAR2 signalling. Cells were harvested at 24 h, and cellular lysates (10 µg protein) were examined for Tau and phospho-Thr181 Tau by western blot analysis. All values were normalised against the respective GAPDH and for comparison, all ratios were calculated against the average from the non-treated cells ± the calculated standard deviation. The number of experiments is shown in each column, and all data groups were determined to have normal distributions and are shown in each column. A SH-SY5Y protein electrophoresis, B SH-SY5Y phospho-Thr181 Tau electrophoresis, C SH-SY5Y Tau protein ratio, and D SH-SY5Y phospho-Thr181 Tau ratio

Article Snippet: Alternatively, the neuronal cells were treated with a rat anti-human antibody (20 μg/ml; AIIB2; Merck KGaA) to block β1-integrin signalling, a mouse anti-human PAR2 antibody, SAM11 (20 μg/ml; Santa Cruz Biotechnology, Heidelberg, Germany), capable of blocking PAR2 signalling, or PAR2-activating peptide (PAR2-AP; 20 μM) to induce PAR2 signalling.

Techniques: Expressing, Phospho-proteomics, Recombinant, Incubation, Blocking Assay, Activity Assay, Control, Western Blot, Comparison, Standard Deviation, Protein Electrophoresis, Electrophoresis

Journal: Cellular and Molecular Neurobiology

Article Title: Procoagulant Extracellular Vesicles Increase Neuronal Tau expression, Metabolism and Processing Through Tissue Factor and Protease Activated Receptor 2

doi: 10.1007/s10571-025-01658-7

Figure Lengend Snippet: Time-course analysis of the Tau protein fragments in differentiated SH-SY5Y cells, following treatment with TF . SH-SY5Y (2 × 10 5 ) were treated with as single dose of recombinant relipidated Innovin TF (0.65 ng/ml) together with human fVIIa (5 nM). In some experiments, the TF aliquots were pre-incubated for 1 h, with 10H10 antibody (20 µg/ml) capable of blocking TF signalling, HTF-1 antibody (20 µg/ml) to block TF-fVIIa protease/procoagulant activity, or a mouse control isotype IgG antibody (20 µg/ml; not shown). Alternatively, the neuronal cells were treated with AIIB2 antibody (20 µg/ml) to block β1-integrin signalling, or SAM11 antibody (20 µg/ml) capable of blocking PAR2 signalling. Sets of cells were harvested at A 48 h and D at 72 h and cellular lysates (10 µg protein) were examined for Tau by western blot analysis. All values were normalised against the respective GAPDH (see Supplementary Fig. 6 A and B) and for comparison, all ratios were calculated against the average from the non-treated cells ± the calculated standard deviation. The data were obtained from 6 biological experiments, and all data groups were determined to have normal distributions. A Electrophoresis at 48 h, B calculated ratios of 50 kDa bands at 48 h, C calculated ratios of 30–35 kDa bands at 48 h, D electrophoresis at 72 h, E calculated ratios of 50 kDa bands at 72 h, F calculated ratios of 40 kDa bands at 72 h, G calculated ratios of 30–35 kDa bands at 72 h

Article Snippet: Alternatively, the neuronal cells were treated with a rat anti-human antibody (20 μg/ml; AIIB2; Merck KGaA) to block β1-integrin signalling, a mouse anti-human PAR2 antibody, SAM11 (20 μg/ml; Santa Cruz Biotechnology, Heidelberg, Germany), capable of blocking PAR2 signalling, or PAR2-activating peptide (PAR2-AP; 20 μM) to induce PAR2 signalling.

Techniques: Recombinant, Incubation, Blocking Assay, Activity Assay, Control, Western Blot, Comparison, Standard Deviation, Electrophoresis

Journal: Particle and Fibre Toxicology

Article Title: Inhibition of protease-activated receptor-2 attenuates multi-walled carbon nanotube exacerbation of allergic lung disease in mice

doi: 10.1186/s12989-025-00648-3

Figure Lengend Snippet: The effect of SAM-11 treatment on allergic lung disease in mice exposed to MWCNTs, HDME, or both. A Representative images of H&E-stained lung sections. B , C Inflammation scores were derived from H&E-stained sections from treated mice. Each data point represents the average of scores from three blinded independent observers. Black arrows indicate inflammation. D Representative images of Masson’s trichrome-stained lung sections showing blue-stained collagen (black arrows). E , F Quantification of trichrome-positive airway collagen. G Representative images of AB-PAS-stained lung tissue sections positive for PAS + mucins stained in purple (black arrows). H , I Morphometric quantification of AB-PAS + airway mucin. Red arrows indicate MWCNTs. Black bars= 100 μm. * p < 0.05, ** p < 0.01, *** p < 0.00, **** p < 0.0001 compared to vehicle; # p < 0.05, #### p < 0.0001 compared to MWCNT; ^^^ p < 0.001, ^^^^ p < 0.0001 compared to HDME; a p <0.05, aaaa p <0.0001 between control and SAM-11 treated mice determined by two-way ANOVA with Tukey’s post-hoc test. Data are presented as mean ± SEM; each data point is an individual mouse. Data for IgG2a isotype control treatment were obtained from a separate independent experiment

Article Snippet: SAM-11 (#sc-13504) and IgG2a (#sc-3878) were purchased from Santa Cruz Biotechnology Inc.

Techniques: Staining, Derivative Assay, Control