rabbit anti mmp 12  (Proteintech)

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