fluorogenic mmp activity assay  (R&D Systems)

Journal: Journal of Nuclear Medicine

Article Title: Preclinical Evaluation of RYM1, a Matrix Metalloproteinase–Targeted Tracer for Imaging Aneurysm

doi: 10.2967/jnumed.116.188656

Figure Lengend Snippet: 99mTc-RYM1 imaging of AAA. (A and B) Examples of fused 99mTc-RYM1 SPECT/CT images of animals from the low remodeling (A) and aneurysm (B) groups, classified on the basis of visual in situ analysis of abdominal aorta. Transversal (left), coronal (middle), and sagittal (right) views are shown. Arrows point to areas of maximal tracer uptake in abdominal aorta. (C) Quantification of 99mTc-RYM1 signal in area of maximal tracer uptake in suprarenal abdominal aorta in low remodeling and AAA groups. *P < 0.05. (D) Correlation between 99mTc-RYM1 signal in vivo and MMP activity quantified by zymography ex vivo. AU = arbitrary units; cpv = counts per voxel; LR = low remodeling.

Article Snippet: RYM1 inhibition constants (K i ) for recombinant human MMP-2, -7, -9, -12, and -13 (R&D Systems) were evaluated in vitro using a fluorogenic MMP activity assay based on Mca-KPLGL-Dpa-AR-NH2 fluorogenic peptide substrate (R&D Systems), as previously described with minor modifications ( 8 ).

Techniques: Imaging, Single Photon Emission Computed Tomography, In Situ, In Vivo, Activity Assay, Zymography, Ex Vivo

Journal: Journal of Nuclear Medicine

Article Title: Preclinical Evaluation of RYM1, a Matrix Metalloproteinase–Targeted Tracer for Imaging Aneurysm

doi: 10.2967/jnumed.116.188656

Figure Lengend Snippet: Ex vivo characterization of suprarenal abdominal aorta. (A) Maximal external diameter of abdominal aorta in low remodeling and AAA groups. (B) Aortic MMP activity quantified by zymography in low remodeling and AAA groups. **P < 0.01. AU = arbitrary units; LR = low remodeling.

Article Snippet: RYM1 inhibition constants (K i ) for recombinant human MMP-2, -7, -9, -12, and -13 (R&D Systems) were evaluated in vitro using a fluorogenic MMP activity assay based on Mca-KPLGL-Dpa-AR-NH2 fluorogenic peptide substrate (R&D Systems), as previously described with minor modifications ( 8 ).

Techniques: Ex Vivo, Activity Assay, Zymography

Journal: Cells

Article Title: High Glucose and Advanced Glycation End Products Induce CD147-Mediated MMP Activity in Human Adipocytes

doi: 10.3390/cells10082098

Figure Lengend Snippet: Effects of HG and CML on MMP2 and MMP9 expression and activity in differentiated adipocytes. Cells were treated with HG (25 mM) or CML (2 µg/mL) with and without NFκB inhibitory peptide (15 µg/mL) for 24 h. ( A ) Quantitative assessment of MMP2 and MMP9 mRNA levels normalized to the house keeping gene, GAPDH using real-time PCR. ( B ) Western blot analysis and quantification of the normalized signal intensity of MMP2 and MMP9 proteins. ( C ) Measurements of soluble MMP2 and MMP9 proteins in cell culture media using corresponding Quantikine ELISA assays. ( D ) Quantitative assessment of MMP2/9 activity using InnoZyme™ Gelatinase (MMP2/MMP9) Fluorogenic Activity Assay. Results represent the means ± SD for three independent experiments. (* p < 0.05) for MMP2 comparison with the corresponding control (HG vs. control; CML vs. BSA; NFκB+HG vs. HG; and NFκB + CML vs. CML). († p < 0.05) for MMP9 comparison with the corresponding control.

Article Snippet: InnoZymeTM Gelatinase (MMP-2/MMP-9) Fluorogenic Activity Assay kit (Millipore Sigma, Burlington, MA, USA) was used to assess the activity of MMPs2 and MMP9 in cell culture media.

Techniques: Expressing, Activity Assay, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Comparison, Control