Journal: BMB Reports

Article Title: SPINK1 promotes cell growth and metastasis of lung adenocarcinoma and acts as a novel prognostic biomarker

doi: 10.5483/BMBRep.2018.51.12.205

Figure Lengend Snippet: SPINK1 enhances the migration and invasion of LAC cells by up-regulating MMP12. (A) MMPs expressions were analyzed by qRT-PCR in A549 cells and H1975 cells transfected with scramble or SPINK1 siRNAs. (B) MMP12 protein expressions were analyzed by Western blot in A549 cells and H1975 cells transfected with scramble or SPINK1 siRNA 1. (C) MMP12 protein levels were measured by Western blot in PC9 cells and H1299 cells transfected with control or SPINK1. (D) MMP12 mRNA expression in xenograft tissues was measured by qPCR. (E) The correlation between SPINK1 expression and MMP12 expression in LAC tissues was analyzed with Pearson correlation. (F) The interference effect of MMP12 siRNA was verified by Western blot in PC9 and H1299 cells stably overexpressing SPINK1. The number of migrating and invasive cells was counted (G). (H) Recombinant human MMP12 (rhMMP12) or control IgG (cIgG) at a concentration of 50 ng/ml was added back into SPINK1 knockdown A549 and H1299 cells, and the number of migrating and invasive cells was counted. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Recombinant human MMP12 (rhMMP12, 917-MPB) and the control IgG (cIgG, 1-001-A) were purchased from R&D Systems.

Techniques: Migration, Quantitative RT-PCR, Transfection, Western Blot, Control, Expressing, Stable Transfection, Recombinant, Concentration Assay, Knockdown

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A comparison of commercially available MMP-12 antibodies as capture antibodies in the FRET activity assay format .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Comparison, Activity Assay

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A plot of RFU (abscissa) vs. analyte (ordinate) in the capture FRET format using antibodies Dendritics 282 (cat# DDX0282), Abcam 11614, or Genway 066 (cat# 15-288-066A) . Analytes are present at 1 ug/mL, except for 12a - autocatalyzed R&D Systems MMP-12 at 1 ng/mL, 12 Wyeth cat - recombinant catalytic MMP-12 at 10 ng/mL, 12 pro - pro form MMP-12, R&D Systems and 12 autocat - autocatalyzed R&D Systems MMP-12 both at 10 ng/mL. n171 and n168 are two sputum samples with unknown MMP-12 concentrations.

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Recombinant

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: Vmax (abscissa) is plotted against Anaspec substrate (ordinate) in a solution phase assay using 20 μM substrate and 100 ng/mL pro or autocatalyzed MMP-12 .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques:

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A pairwise plot of interpolated MMP-12 concentrations for both activity and total MMP-12 protein concentration in normal, COPD, and asthmatic donors .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Activity Assay, Protein Concentration

Journal: BMC Pulmonary Medicine

Article Title: Optimization of total protein and activity assays for the detection of MMP-12 in induced human sputum

doi: 10.1186/1471-2466-10-40

Figure Lengend Snippet: A boxplot of condition (abscissa) vs interpolated MMP-12 protein concentration (ordinate) trellised by assay format .

Article Snippet: MMP-12 protein from R&D Systems was autocatalyzed and spiked into four human sputum samples and into assay diluent at three different concentrations (final theoretical spiking concentrations of 10,000, 3000, and 1000 pg/mL).

Techniques: Protein Concentration