anti mmp 12  (R&D Systems)

Journal: iScience

Article Title: Human monocyte-derived macrophages shift subcellular metalloprotease activity depending on their activation state

doi: 10.1016/j.isci.2024.111171

Figure Lengend Snippet:

Article Snippet: After protein transfer, membranes were blocked for 5 min in EveryBlot Blocking buffer (Cat # 12010020, Biorad) and incubated with anti-MMP-9 (Cat # AB911, R&D systems), anti-MMP-12 (Cat # AF917, R&D systems), anti-ADAM-9 (Cat # DY939, Detection antibody of Human DuoSet ELISA, R&D systems), anti-SP1 (Cat # 5931, Cell Signaling), anti-β-actin (Cat # 20536-1, Proteintech), anti-H3 (Cat # 9715, Cell Signaling), anti-CD44 (Cat # 3570, Cell Signaling), anti-HSP90 (Cat # MAB3286, R&D systems), anti-TIMP-1 (Cat # 8946S, Cell Signaling) or anti-A2M (Cat # AF1938, R&D Systems) antibodies diluted in 10% EveryBlot Blocking buffer in TBS-T (1× TBS containing 0.1% Tween 20) overnight at 4°C.

Techniques: Plasmid Preparation, Recombinant, Knock-Out, Blocking Assay, Staining, Antibody Labeling, Reverse Transcription, Lysis, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Fractionation, Cell Culture, Software, Flow Cytometry, Fluorescence, Spectrophotometry

Journal: iScience

Article Title: Human monocyte-derived macrophages shift subcellular metalloprotease activity depending on their activation state

doi: 10.1016/j.isci.2024.111171

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Article Snippet: Anti-Human MMP-12 , R&D systems , Cat # AF917; RRID: AB_3644613.

Techniques: Plasmid Preparation, Recombinant, Knock-Out, Blocking Assay, Staining, Antibody Labeling, Reverse Transcription, Lysis, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Fractionation, Cell Culture, Software, Flow Cytometry, Fluorescence, Spectrophotometry

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Expression of MMP-12 mRNA by in-situ ASMC. Laser capture microdissection was performed to collect ASMC from bronchial biopsy sections (A). MMP-12 mRNA expression was analysed by real-time RT-PCR (B). The data, obtained from 4 normal volunteers, 3 asthma, 3 COPD and 5 chronic cough patients, were normalized to the housekeeping gene 18S rRNA representative of relative MMP-12 mRNA expression (C). Lung tissue was used as a positive control.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Expressing, In Situ, Laser Capture Microdissection, Quantitative RT-PCR, Positive Control

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Expression of MMP-12 protein in bronchial smooth muscle tissue and cell cultures by immunostaining. Sections from human bronchial samples were prepared (A). Cell cultures were incubated on 8-well chamber slides for 72 hours in the absence (B) or presence (C) of 10 ng/ml IL-1β. Immunostaining was performed to detect MMP-12 expression using a rabbit anti-MMP-12 antibody. The primary antibody was replaced by a normal rabbit immunoglobulin as a negative control (D). Bar = 50 mm. Results are representative from three donors.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Expressing, Immunostaining, Incubation, Negative Control

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Stimulation of MMP-12 mRNA expression by IL-1β in ASMC. Cells were incubated in the absence or presence of IL-1β at 10 ng/ml for 1–24 hours (A, B), or for 24 hours over 0.01 to 10 ng/ml (C). MMP-12 and GAPDH mRNA expression was analysed by RT-PCR (A) or real-time RT-PCR (B,C). Results (B,C) were expressed as a ratio of target gene to GAPDH mRNA control and are the mean ± SEM from three ASMC donors. **P < 0.01 compared with control.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Expressing, Incubation, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Western blot analysis of MMP-12 protein expression in ASMC and the effect of IL-1β. Cells were incubated in 6-well plates in the absence or presence of 10 ng/ml IL-1β for 24–72 hours. MMP-12 protein expression was analysed by Western blot (A) and shown as a ratio of GAPDH obtained by densitometric analysis (B). Results are mean ± SEM from three ASMC donors.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Western Blot, Expressing, Incubation

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Activity of MMP-12 secreted by ASMC and stimulation by IL-1β. (A) Gelatin zymography detection of activity of MMPs released into the conditioned media by ASMC treated for 48 hours (lane 1, protein markers; lane 2, control; lane 3, 10 ng/ml IL-1β; lane 4, 10 ng/ml TGF-β; lane 5, TGF-β plus IL-1β; lane 6, 10 ng/ml TNF-α; lane 7, TNF-α plus IL-1β; lane 8, 10 ng/ml IL-13). (B) Time-dependent stimulation of MMP-12 activity by 10 ng/ml IL-1β. Lane s, human MMP-12 standard protein used as a positive control. Lane n, negative control. (C) Concentration-dependent stimulation of MMP-12 activity by IL-1β for 48 hours. Relevant band intensities were quantified by scanning densitometric analysis and normalized to 5 × 10 5 cells. Results are the mean ± SEM from three ASMC donors. *P < 0.05, **P < 0.01 compared with control.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Activity Assay, Zymography, Positive Control, Negative Control, Concentration Assay

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Inhibition of IL-1β-stimulated MMP-12 activity and mRNA expression by PD98059 and SP600125. ASMC were pre-treated for 1 hour with a specific inhibitor for ERK, PD98059 (1 or 10 μM) or for JNK, SP600125 (10 μM), and then were co-treated with 10 ng/ml IL-1β. (A) MMP-12 activity in conditioned media was detected after 48 hours by zymography. The relevant band intensities were quantified by scanning densitometric analysis. (B) MMP-12 mRNA was analysed after 24 hours by real-time RT-PCR. The data are expressed as the percentage of IL-1β alone and are the mean ± SEM from three ASMC donors. *P < 0.05, **P < 0.01 compared with IL-1β alone.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Inhibition, Activity Assay, Expressing, Zymography, Quantitative RT-PCR

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Inhibition of IL-1β-stimulated MMP-12 activity and mRNA expression by wortmannin, LY294002 and dexamethasone. ASMC were pre-treated for 1 hour with PI3-K inhibitors, wortmannin or LY294002, or dexamethasone at the indicated concentrations, and then were co-treated with 10 ng/ml IL-1β. (A) MMP-12 activity in conditioned media was detected after 48 hours by zymography. The relevant band intensities were quantified by scanning densitometric analysis. (B) MMP-12 mRNA was analysed after 24 hours by real-time RT-PCR. The data are expressed as the percentage of IL-1β alone and are the mean ± SEM from three ASMC donors. *P < 0.05, **P < 0.01 compared with IL-1β alone.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Inhibition, Activity Assay, Expressing, Zymography, Quantitative RT-PCR

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Effects of TNF-α and TGF-β1 on IL-1β-stimulated MMP-12 activity, mRNA expression and c-Jun activation. ASMC were treated with 10 ng/ml of TNF-α or TGF-β1 alone, or in combination with 10 ng/ml IL-1β. (A) MMP-12 activity in condition media was detected by zymography after 48 hours, and the relevant band intensities were quantified by densitometric analysis and normalized to 5 × 10 5 cells. (B) MMP-12 mRNA expression was determined by real-time RT-PCR after 24 hours and expressed as a ratio to GAPDH mRNA. (C) c-Jun activation and DNA-binding analysed by the TransAM AP-1 family kit. Data are the mean ± SEM from three ASMC donors. *P < 0.05, **P < 0.01 compared with control; +P < 0.05, ++P < 0.01, compared with IL-1β alone.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Activity Assay, Expressing, Activation Assay, Zymography, Quantitative RT-PCR, Binding Assay

Journal: Respiratory Research

Article Title: Induction and regulation of matrix metalloproteinase-12in human airway smooth muscle cells

doi: 10.1186/1465-9921-6-148

Figure Lengend Snippet: Western blot analysis of MMP-12 secretion by ASMC. Cells were incubated for 48 hours with 10 ng/ml IL-1β or TNF-α alone, or IL-1β combination with TNF-α, PD98059 (1 μM), SP600125 (10 μM) or wortmannin (250 nM) for 48 hours. MMP-12 protein released into conditioned media was determined by Western blot using 20 μl of 40-fold concentrated samples. Relevant band intensities were quantified by scanning densitometric analysis and normalized to 5 × 10 5 cells. Results are the mean ± SEM from three ASMC donors. **P < 0.01 compared with control; +P < 0.05, +P < 0.01 compared with IL-1β alone.

Article Snippet: Rabbit anti-human MMP-12 antibodies (AB19053 and AB19051) were obtained from Chemicon (Hampshire, UK).

Techniques: Western Blot, Incubation

Journal: Journal of Clinical Medicine

Article Title: Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing

doi: 10.3390/jcm9113458

Figure Lengend Snippet: List of DEGs when the IgG4-ROD tissue was compared to all control tissues.

Article Snippet: Immunostaining by mouse anti-human MMP12 (1:50, MAB919, R&D Systems, Minneapolis, MN, USA) and rabbit anti-human SPP1 (1:1000, HPA027541, Atlas Antibodies, Bromma, Sweden) was performed using paraffin-embedded biopsy tissues.

Techniques: Control

Journal: Journal of Clinical Medicine

Article Title: Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing

doi: 10.3390/jcm9113458

Figure Lengend Snippet: Differentially expressed genes of IgG4-ROD versus orbital MALT lymphoma: top 40 in descending p value.

Article Snippet: Immunostaining by mouse anti-human MMP12 (1:50, MAB919, R&D Systems, Minneapolis, MN, USA) and rabbit anti-human SPP1 (1:1000, HPA027541, Atlas Antibodies, Bromma, Sweden) was performed using paraffin-embedded biopsy tissues.

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing

doi: 10.3390/jcm9113458

Figure Lengend Snippet: Differentially expressed genes of IgG4-ROD versus RLH: top 40 in descending p value.

Article Snippet: Immunostaining by mouse anti-human MMP12 (1:50, MAB919, R&D Systems, Minneapolis, MN, USA) and rabbit anti-human SPP1 (1:1000, HPA027541, Atlas Antibodies, Bromma, Sweden) was performed using paraffin-embedded biopsy tissues.

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing

doi: 10.3390/jcm9113458

Figure Lengend Snippet: Results of real-time PCR for validation of differentially expressed genes obtained from RNA sequencing. *** Messenger RNA of MMP12 and SPP1 in immunoglobulin G4-related ophthalmic disease is significantly higher than mucosa-associated lymphoid tissue lymphoma ( p = 2.7141 × 10 −8 and 1.2044 × 10 −7 , respectively). Statistical analysis was performed by a Mann–Whitney U test ( n = 30). IgG4-ROD: Immunoglobulin G4 related ophthalmic disease, MALT: Mucosa-associated lymph tissue, MMP12: Matrix metallopeptidase 12, SPP1: Secreted phosphoprotein 1.

Article Snippet: Immunostaining by mouse anti-human MMP12 (1:50, MAB919, R&D Systems, Minneapolis, MN, USA) and rabbit anti-human SPP1 (1:1000, HPA027541, Atlas Antibodies, Bromma, Sweden) was performed using paraffin-embedded biopsy tissues.

Techniques: Real-time Polymerase Chain Reaction, Biomarker Discovery, RNA Sequencing, MANN-WHITNEY

Journal: Journal of Clinical Medicine

Article Title: Comprehensive Gene Analysis of IgG4-Related Ophthalmic Disease Using RNA Sequencing

doi: 10.3390/jcm9113458

Figure Lengend Snippet: Representative immunohistochemical staining patterns of formalin-fixed, paraffin-embedded sections (MMP12: magnification 400×, bar: 50 μm, SPP1: magnification 200×, bar: 100μm). The upper row is immunoreactivity of MMP12, and the lower row is immunoreactivity of SPP1. From left column to right column, IgG4-ROD, MALT lymphoma, RLH, and the lacrimal gland tissue section are shown. Many MMP12 positive cells are observable in the fibrotic areas and follicular areas of IgG4-ROD tissues. SPP1 is intensely stained in the fibrotic part of IgG4-ROD. IgG4-ROD: Immunoglobulin G4 related ophthalmic disease, MALT: Mucosa-associated lymph tissue, RLH: Reactive lymphoid hyperplasia, MMP12: Matrix metallopeptidase 12, SPP1: Secreted phosphoprotein 1.

Article Snippet: Immunostaining by mouse anti-human MMP12 (1:50, MAB919, R&D Systems, Minneapolis, MN, USA) and rabbit anti-human SPP1 (1:1000, HPA027541, Atlas Antibodies, Bromma, Sweden) was performed using paraffin-embedded biopsy tissues.

Techniques: Immunohistochemical staining, Staining, Formalin-fixed Paraffin-Embedded