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par2 activating matriptase  (R&D Systems)


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    R&D Systems par2 activating matriptase
    Colonic <t>PAR2</t> activation by 2F elicits LSN responses that are mediated by endosomal internalization, PKA and PKC. (A) Experimental framework for LSN recordings showing a cannulated colon in a recording chamber with the LSN aspirated into a suction electrode. (B) Representative action potentials recorded from LSN afferent fiber prestimulation. (C and D) 100 µM 2F application elicits LSN responses illustrated by (C) action potential traces prestimulation and 15 minutes poststimulation and (D) change in LSN firing rate over time after stimulation via the luminal inflow (vertical dotted line). 2F-stimulation increased LSN firing (E and F). Timeline and peak change in firing rate after 100 µM 2F application (vertical dotted line) in tissue pretreated with (E) 50 µM PitStop2 (PS2) inhibitor for endosomal internalization or negative control PitNot2 (PN2) and (F) 100 µM H-89 dihydrochloride (H-89) and bisindolylmaleimide (GFX), PKA and PKC inhibitors or DMSO vehicle. (E) PS2 as well as separate and simultaneous pretreatment with H-89 and GFX reduced the peak response to 2F. (D and E) Independent samples t test comparing the peak change in firing rate (N = 5–9). (F) One-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–6). * P < 0.05, ** P < 0.01, *** P < 0.001 **** P < 0.0001. Data are presented as mean ± SD. LSN, lumbar splanchnic nerve; PAR2, <t>protease-activated</t> <t>receptor</t> <t>2;</t> PKA, protein kinase A; PKC, protein kinase C.
    Par2 Activating Matriptase, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/human+matriptase/Recombinant+Human+Matriptase%2FST14+Catalytic+Domain%2C+CF/pmc13098787-140-26-30
    Average 95 stars, based on 30 article reviews
    par2 activating matriptase - by Bioz Stars, 2026-09
    95/100 stars

    Images

    1) Product Images from "Monoclonal antibody inhibition of PAR2 reduces phenotype severity and pain in murine inflammatory bowel disease"

    Article Title: Monoclonal antibody inhibition of PAR2 reduces phenotype severity and pain in murine inflammatory bowel disease

    Journal: Pain Reports

    doi: 10.1097/PR9.0000000000001446

    Colonic PAR2 activation by 2F elicits LSN responses that are mediated by endosomal internalization, PKA and PKC. (A) Experimental framework for LSN recordings showing a cannulated colon in a recording chamber with the LSN aspirated into a suction electrode. (B) Representative action potentials recorded from LSN afferent fiber prestimulation. (C and D) 100 µM 2F application elicits LSN responses illustrated by (C) action potential traces prestimulation and 15 minutes poststimulation and (D) change in LSN firing rate over time after stimulation via the luminal inflow (vertical dotted line). 2F-stimulation increased LSN firing (E and F). Timeline and peak change in firing rate after 100 µM 2F application (vertical dotted line) in tissue pretreated with (E) 50 µM PitStop2 (PS2) inhibitor for endosomal internalization or negative control PitNot2 (PN2) and (F) 100 µM H-89 dihydrochloride (H-89) and bisindolylmaleimide (GFX), PKA and PKC inhibitors or DMSO vehicle. (E) PS2 as well as separate and simultaneous pretreatment with H-89 and GFX reduced the peak response to 2F. (D and E) Independent samples t test comparing the peak change in firing rate (N = 5–9). (F) One-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–6). * P < 0.05, ** P < 0.01, *** P < 0.001 **** P < 0.0001. Data are presented as mean ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.
    Figure Legend Snippet: Colonic PAR2 activation by 2F elicits LSN responses that are mediated by endosomal internalization, PKA and PKC. (A) Experimental framework for LSN recordings showing a cannulated colon in a recording chamber with the LSN aspirated into a suction electrode. (B) Representative action potentials recorded from LSN afferent fiber prestimulation. (C and D) 100 µM 2F application elicits LSN responses illustrated by (C) action potential traces prestimulation and 15 minutes poststimulation and (D) change in LSN firing rate over time after stimulation via the luminal inflow (vertical dotted line). 2F-stimulation increased LSN firing (E and F). Timeline and peak change in firing rate after 100 µM 2F application (vertical dotted line) in tissue pretreated with (E) 50 µM PitStop2 (PS2) inhibitor for endosomal internalization or negative control PitNot2 (PN2) and (F) 100 µM H-89 dihydrochloride (H-89) and bisindolylmaleimide (GFX), PKA and PKC inhibitors or DMSO vehicle. (E) PS2 as well as separate and simultaneous pretreatment with H-89 and GFX reduced the peak response to 2F. (D and E) Independent samples t test comparing the peak change in firing rate (N = 5–9). (F) One-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–6). * P < 0.05, ** P < 0.01, *** P < 0.001 **** P < 0.0001. Data are presented as mean ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.

    Techniques Used: Activation Assay, Negative Control

    PAR2 activation in the colon by 2F sensitizes LSN responses to mechanical and chemical stimulation of the colon via endosomal internalization, PKA and PKC. LSN action potential firing after mechanical distention: gradually increasing intraluminal pressure from 0 to 80 mm Hg, and chemical application: 1 mM cinnamaldehyde and 1 µM capsaicin are illustrated in (A). (B) Responses to distention were quantified through the change in firing rate over increasing discrete pressure values. (C and D) Responses to chemical stimuli were quantified through the change in firing rate over time after application (vertical dotted line). 100 µM 2F stimulation, but not vehicle control, elicited sensitization of the LSN to (B) distention, (C) cinnamaldehyde, and (D) capsaicin. Pretreatment with PS2 before 2F stimulation reduced subsequent LSN responses to distention, cinnamaldehyde, and capsaicin compared with PN2. Pretreatment with H-89 and GFX, applied either simultaneously or individually, also reduced subsequent peak LSN responses to distention, cinnamaldehyde, and capsaicin compared with DMSO vehicle. (B–D) Independent samples t tests and one-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–9). * P < 0.05, ** P < 0.01, **** P < 0.0001. Data are presented as means ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.
    Figure Legend Snippet: PAR2 activation in the colon by 2F sensitizes LSN responses to mechanical and chemical stimulation of the colon via endosomal internalization, PKA and PKC. LSN action potential firing after mechanical distention: gradually increasing intraluminal pressure from 0 to 80 mm Hg, and chemical application: 1 mM cinnamaldehyde and 1 µM capsaicin are illustrated in (A). (B) Responses to distention were quantified through the change in firing rate over increasing discrete pressure values. (C and D) Responses to chemical stimuli were quantified through the change in firing rate over time after application (vertical dotted line). 100 µM 2F stimulation, but not vehicle control, elicited sensitization of the LSN to (B) distention, (C) cinnamaldehyde, and (D) capsaicin. Pretreatment with PS2 before 2F stimulation reduced subsequent LSN responses to distention, cinnamaldehyde, and capsaicin compared with PN2. Pretreatment with H-89 and GFX, applied either simultaneously or individually, also reduced subsequent peak LSN responses to distention, cinnamaldehyde, and capsaicin compared with DMSO vehicle. (B–D) Independent samples t tests and one-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–9). * P < 0.05, ** P < 0.01, **** P < 0.0001. Data are presented as means ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.

    Techniques Used: Activation Assay, Control

    Related Articles

    Infection:

    Article Title: SARS-CoV-2 infection of human lung epithelial cells induces TMPRSS-mediated acute fibrin deposition.
    Article Snippet: .. The cleavage of Thrb-324 peptide was initiated by mixing 10 μM of the peptide with 100 ng of human factor Xa (R&D Systems, Inc), or 400ng of human matriptase (R&D Systems, Inc) in 100 μl assay buffer containing 25mMTris at pH9.0, 2.5μMZnCl2, and0.005%Brij-35 (w/v), or with infected cells or 100 μl of infected supernatant in 96-well plates. ..

    Article Title: SARS-CoV-2 infection of human lung epithelial cells induces TMPRSS-mediated acute fibrin deposition
    Article Snippet: .. The cleavage of Thrb-324 peptide was initiated by mixing 10 μM of the peptide with 100 ng of human factor Xa (R&D Systems, Inc), or 400 ng of human matriptase (R&D Systems, Inc) in 100 μl assay buffer containing 25 mM Tris at pH 9.0, 2.5 μM ZnCl 2 , and 0.005% Brij-35 (w/v), or with infected cells or 100 μl of infected supernatant in 96-well plates. ..

    Purification:

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: .. Cleavage of the Prot-mαCD3 IgG was performed by addition of 1l of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) at a concentration of approx. ..

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: Cytokine (IFN-γ, TNF-α, IL-2, GM-CSF) and cytotoxic granule (granzyme B) secretion was assessed 48 h after incubation of target cells with TCB and PBMCs (as above) using Human Soluble Protein Master Buffer Kit (BD Biosciences) according to the manufacturer’s protocol. .. The cleavage of the Prot-FOLR1-TCB was done by incubation of 1 μl of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) with approx. ..

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: .. 2 h after the addition of PBMCs and TCBs is indicated as time 0 h. The cleavage of the Prot-FOLR1-TCB was done by incubation of 1 μl of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) with approx. ..

    Recombinant:

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: .. Cleavage of the Prot-mαCD3 IgG was performed by addition of 1l of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) at a concentration of approx. ..

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: Cytokine (IFN-γ, TNF-α, IL-2, GM-CSF) and cytotoxic granule (granzyme B) secretion was assessed 48 h after incubation of target cells with TCB and PBMCs (as above) using Human Soluble Protein Master Buffer Kit (BD Biosciences) according to the manufacturer’s protocol. .. The cleavage of the Prot-FOLR1-TCB was done by incubation of 1 μl of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) with approx. ..

    Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer
    Article Snippet: The reaction was monitored by measuring the absorbance at 405 nm every 30 s for 30 min using the SpectraMax M5 plate reader (Molecular Devices). .. In matriptase inhibition assay, 50 μL of recombinant human matriptase (R&D Systems, Minneapolis, MN, USA; 3946-SE-010) (3.8 mM prepared in assay buffer: 50 mM Tris, 50 mM NaCl, 0.01% Tween 20; pH 9.0) and 1 μL of different concentrations of inhibitors ( 2 and 3 ; half log dilutions prepared in DMSO) or the solvent control were pre-incubated at room temperature for 15 min. .. Following the incubation period, 50 μL of the 50 μM substrate solution (Boc-Gln-Ala-Arg-AMC) (R&D Systems, Minneapolis, MN, USA; ES014) were added to each well, and the enzyme activity was monitored by measuring the increase in fluorescence signal from the fluorescently-labeled substrate every 30 s for 30 min (Ex 380 nm, Em 460 nm) using the SpectraMax M5 plate reader (Molecular Devices).

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: .. 2 h after the addition of PBMCs and TCBs is indicated as time 0 h. The cleavage of the Prot-FOLR1-TCB was done by incubation of 1 μl of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) with approx. ..

    Concentration Assay:

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: .. Cleavage of the Prot-mαCD3 IgG was performed by addition of 1l of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) at a concentration of approx. ..

    Incubation:

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: Cytokine (IFN-γ, TNF-α, IL-2, GM-CSF) and cytotoxic granule (granzyme B) secretion was assessed 48 h after incubation of target cells with TCB and PBMCs (as above) using Human Soluble Protein Master Buffer Kit (BD Biosciences) according to the manufacturer’s protocol. .. The cleavage of the Prot-FOLR1-TCB was done by incubation of 1 μl of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) with approx. ..

    Article Title: Design and Characterization of Prodrugged Anti‐CTLA‐4 Antibodies
    Article Snippet: .. Prodrugged antibody was incubated with human matriptase (30:1 molar ratio, R&D system, 3946‐SE‐010) in 100 mM Tris buffer, pH7.6 at 37C. ..

    Article Title: Design and Characterization of Prodrugged Anti-CTLA-4 Antibodies.
    Article Snippet: Therapeutic antibodies are widely used to treat diseases like cancer and inflammatory conditions by binding with high specificity to their molecular targets.. Masking is a strategy to mitigate undesirable 10.1002/cbic.202500304 A cc ep te d M an us cr ip t ChemBioChem This article is protected by copyright.. All rights reserved.

    Article Title: Protease-activation using anti-idiotypic masks enables tumor specificity of a folate receptor 1-T cell bispecific antibody
    Article Snippet: .. 2 h after the addition of PBMCs and TCBs is indicated as time 0 h. The cleavage of the Prot-FOLR1-TCB was done by incubation of 1 μl of purified recombinant human matriptase (0.44 mg/ml, R&D Systems) with approx. ..

    Inhibition:

    Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer
    Article Snippet: The reaction was monitored by measuring the absorbance at 405 nm every 30 s for 30 min using the SpectraMax M5 plate reader (Molecular Devices). .. In matriptase inhibition assay, 50 μL of recombinant human matriptase (R&D Systems, Minneapolis, MN, USA; 3946-SE-010) (3.8 mM prepared in assay buffer: 50 mM Tris, 50 mM NaCl, 0.01% Tween 20; pH 9.0) and 1 μL of different concentrations of inhibitors ( 2 and 3 ; half log dilutions prepared in DMSO) or the solvent control were pre-incubated at room temperature for 15 min. .. Following the incubation period, 50 μL of the 50 μM substrate solution (Boc-Gln-Ala-Arg-AMC) (R&D Systems, Minneapolis, MN, USA; ES014) were added to each well, and the enzyme activity was monitored by measuring the increase in fluorescence signal from the fluorescently-labeled substrate every 30 s for 30 min (Ex 380 nm, Em 460 nm) using the SpectraMax M5 plate reader (Molecular Devices).

    Solvent:

    Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer
    Article Snippet: The reaction was monitored by measuring the absorbance at 405 nm every 30 s for 30 min using the SpectraMax M5 plate reader (Molecular Devices). .. In matriptase inhibition assay, 50 μL of recombinant human matriptase (R&D Systems, Minneapolis, MN, USA; 3946-SE-010) (3.8 mM prepared in assay buffer: 50 mM Tris, 50 mM NaCl, 0.01% Tween 20; pH 9.0) and 1 μL of different concentrations of inhibitors ( 2 and 3 ; half log dilutions prepared in DMSO) or the solvent control were pre-incubated at room temperature for 15 min. .. Following the incubation period, 50 μL of the 50 μM substrate solution (Boc-Gln-Ala-Arg-AMC) (R&D Systems, Minneapolis, MN, USA; ES014) were added to each well, and the enzyme activity was monitored by measuring the increase in fluorescence signal from the fluorescently-labeled substrate every 30 s for 30 min (Ex 380 nm, Em 460 nm) using the SpectraMax M5 plate reader (Molecular Devices).

    Control:

    Article Title: Kempopeptin C, a Novel Marine-Derived Serine Protease Inhibitor Targeting Invasive Breast Cancer
    Article Snippet: The reaction was monitored by measuring the absorbance at 405 nm every 30 s for 30 min using the SpectraMax M5 plate reader (Molecular Devices). .. In matriptase inhibition assay, 50 μL of recombinant human matriptase (R&D Systems, Minneapolis, MN, USA; 3946-SE-010) (3.8 mM prepared in assay buffer: 50 mM Tris, 50 mM NaCl, 0.01% Tween 20; pH 9.0) and 1 μL of different concentrations of inhibitors ( 2 and 3 ; half log dilutions prepared in DMSO) or the solvent control were pre-incubated at room temperature for 15 min. .. Following the incubation period, 50 μL of the 50 μM substrate solution (Boc-Gln-Ala-Arg-AMC) (R&D Systems, Minneapolis, MN, USA; ES014) were added to each well, and the enzyme activity was monitored by measuring the increase in fluorescence signal from the fluorescently-labeled substrate every 30 s for 30 min (Ex 380 nm, Em 460 nm) using the SpectraMax M5 plate reader (Molecular Devices).



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    human mt sp1  (R&D Systems)
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    Structures of the masking reagent consist of: 1) a maleimide for conjugation, 2) a <t>matriptase‐cleavable</t> peptide (highlighted in purple), and 3) PEG to create steric hindrance, preventing the antibody from binding to the target.
    Human Mt Sp1, supplied by R&D Systems, 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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    sheep  (R&D Systems)
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    Structures of the masking reagent consist of: 1) a maleimide for conjugation, 2) a <t>matriptase‐cleavable</t> peptide (highlighted in purple), and 3) PEG to create steric hindrance, preventing the antibody from binding to the target.
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    Image Search Results


    Colonic PAR2 activation by 2F elicits LSN responses that are mediated by endosomal internalization, PKA and PKC. (A) Experimental framework for LSN recordings showing a cannulated colon in a recording chamber with the LSN aspirated into a suction electrode. (B) Representative action potentials recorded from LSN afferent fiber prestimulation. (C and D) 100 µM 2F application elicits LSN responses illustrated by (C) action potential traces prestimulation and 15 minutes poststimulation and (D) change in LSN firing rate over time after stimulation via the luminal inflow (vertical dotted line). 2F-stimulation increased LSN firing (E and F). Timeline and peak change in firing rate after 100 µM 2F application (vertical dotted line) in tissue pretreated with (E) 50 µM PitStop2 (PS2) inhibitor for endosomal internalization or negative control PitNot2 (PN2) and (F) 100 µM H-89 dihydrochloride (H-89) and bisindolylmaleimide (GFX), PKA and PKC inhibitors or DMSO vehicle. (E) PS2 as well as separate and simultaneous pretreatment with H-89 and GFX reduced the peak response to 2F. (D and E) Independent samples t test comparing the peak change in firing rate (N = 5–9). (F) One-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–6). * P < 0.05, ** P < 0.01, *** P < 0.001 **** P < 0.0001. Data are presented as mean ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.

    Journal: Pain Reports

    Article Title: Monoclonal antibody inhibition of PAR2 reduces phenotype severity and pain in murine inflammatory bowel disease

    doi: 10.1097/PR9.0000000000001446

    Figure Lengend Snippet: Colonic PAR2 activation by 2F elicits LSN responses that are mediated by endosomal internalization, PKA and PKC. (A) Experimental framework for LSN recordings showing a cannulated colon in a recording chamber with the LSN aspirated into a suction electrode. (B) Representative action potentials recorded from LSN afferent fiber prestimulation. (C and D) 100 µM 2F application elicits LSN responses illustrated by (C) action potential traces prestimulation and 15 minutes poststimulation and (D) change in LSN firing rate over time after stimulation via the luminal inflow (vertical dotted line). 2F-stimulation increased LSN firing (E and F). Timeline and peak change in firing rate after 100 µM 2F application (vertical dotted line) in tissue pretreated with (E) 50 µM PitStop2 (PS2) inhibitor for endosomal internalization or negative control PitNot2 (PN2) and (F) 100 µM H-89 dihydrochloride (H-89) and bisindolylmaleimide (GFX), PKA and PKC inhibitors or DMSO vehicle. (E) PS2 as well as separate and simultaneous pretreatment with H-89 and GFX reduced the peak response to 2F. (D and E) Independent samples t test comparing the peak change in firing rate (N = 5–9). (F) One-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–6). * P < 0.05, ** P < 0.01, *** P < 0.001 **** P < 0.0001. Data are presented as mean ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.

    Article Snippet: When measuring the inhibitory concentration 50 (IC50), cells received PAR650097 mIgG or hIgG or isotype control antibodies, for 60 minutes at room temperature before addition of PAR2 activating matriptase (3946-SEB-010, R&D Systems, Abingdon, United Kingdom).

    Techniques: Activation Assay, Negative Control

    PAR2 activation in the colon by 2F sensitizes LSN responses to mechanical and chemical stimulation of the colon via endosomal internalization, PKA and PKC. LSN action potential firing after mechanical distention: gradually increasing intraluminal pressure from 0 to 80 mm Hg, and chemical application: 1 mM cinnamaldehyde and 1 µM capsaicin are illustrated in (A). (B) Responses to distention were quantified through the change in firing rate over increasing discrete pressure values. (C and D) Responses to chemical stimuli were quantified through the change in firing rate over time after application (vertical dotted line). 100 µM 2F stimulation, but not vehicle control, elicited sensitization of the LSN to (B) distention, (C) cinnamaldehyde, and (D) capsaicin. Pretreatment with PS2 before 2F stimulation reduced subsequent LSN responses to distention, cinnamaldehyde, and capsaicin compared with PN2. Pretreatment with H-89 and GFX, applied either simultaneously or individually, also reduced subsequent peak LSN responses to distention, cinnamaldehyde, and capsaicin compared with DMSO vehicle. (B–D) Independent samples t tests and one-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–9). * P < 0.05, ** P < 0.01, **** P < 0.0001. Data are presented as means ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.

    Journal: Pain Reports

    Article Title: Monoclonal antibody inhibition of PAR2 reduces phenotype severity and pain in murine inflammatory bowel disease

    doi: 10.1097/PR9.0000000000001446

    Figure Lengend Snippet: PAR2 activation in the colon by 2F sensitizes LSN responses to mechanical and chemical stimulation of the colon via endosomal internalization, PKA and PKC. LSN action potential firing after mechanical distention: gradually increasing intraluminal pressure from 0 to 80 mm Hg, and chemical application: 1 mM cinnamaldehyde and 1 µM capsaicin are illustrated in (A). (B) Responses to distention were quantified through the change in firing rate over increasing discrete pressure values. (C and D) Responses to chemical stimuli were quantified through the change in firing rate over time after application (vertical dotted line). 100 µM 2F stimulation, but not vehicle control, elicited sensitization of the LSN to (B) distention, (C) cinnamaldehyde, and (D) capsaicin. Pretreatment with PS2 before 2F stimulation reduced subsequent LSN responses to distention, cinnamaldehyde, and capsaicin compared with PN2. Pretreatment with H-89 and GFX, applied either simultaneously or individually, also reduced subsequent peak LSN responses to distention, cinnamaldehyde, and capsaicin compared with DMSO vehicle. (B–D) Independent samples t tests and one-way ANOVA with post-hoc FDR-corrected independent samples t test (N = 5–9). * P < 0.05, ** P < 0.01, **** P < 0.0001. Data are presented as means ± SD. LSN, lumbar splanchnic nerve; PAR2, protease-activated receptor 2; PKA, protein kinase A; PKC, protein kinase C.

    Article Snippet: When measuring the inhibitory concentration 50 (IC50), cells received PAR650097 mIgG or hIgG or isotype control antibodies, for 60 minutes at room temperature before addition of PAR2 activating matriptase (3946-SEB-010, R&D Systems, Abingdon, United Kingdom).

    Techniques: Activation Assay, Control

    Structures of the masking reagent consist of: 1) a maleimide for conjugation, 2) a matriptase‐cleavable peptide (highlighted in purple), and 3) PEG to create steric hindrance, preventing the antibody from binding to the target.

    Journal: Chembiochem

    Article Title: Design and Characterization of Prodrugged Anti‐CTLA‐4 Antibodies

    doi: 10.1002/cbic.202500304

    Figure Lengend Snippet: Structures of the masking reagent consist of: 1) a maleimide for conjugation, 2) a matriptase‐cleavable peptide (highlighted in purple), and 3) PEG to create steric hindrance, preventing the antibody from binding to the target.

    Article Snippet: Prodrugged antibody was incubated with human matriptase (30:1 molar ratio, R&D system, 3946‐SE‐010) in 100 mM Tris buffer, pH7.6 at 37C.

    Techniques: Conjugation Assay, Binding Assay