mmp 9 elisa  (Boster Bio)

Journal: Cellular microbiology

Article Title: Filifactor alocis modulates human neutrophil antimicrobial functional responses

doi: 10.1111/cmi.12829

Figure Lengend Snippet: Opsonized F. alocis induces exocytosis of three of the four neutrophil granule subtypes Neutrophils were unchallenged (basal), challenged with fMLF (5 min), or challenged with live or heat killed (HK) opsonized F. alocis (MOI 10, 30 min). A-B . Secretory vesicle and specific granule exocytosis was determined via flow cytometry by measuring the increase in plasma membrane expression of CD35 or CD66b respectively. Data are expressed as the mean ± SEM of the mean channel of fluorescence (mcf) from 6 independent experiments. In A , ns: non-significant compared to basal. C . Gelatinase granule exocytosis was measured by ELISA as levels of matrix metalloproteinase 9 (MMP-9) present in the supernatants collected from the different experimental conditions. Data are expressed as mean ± SEM of MMP-9 release in ng/4 × 10 6 cells, from 5 independent experiments. D. Gelatin zymography gels were used to determine the activity of MMP-9 in the supernatants from the same experimental conditions described above. The protein standard is shown on the left side, and the different MMP-9 isoforms are also indicated. E. Quantification of the densitometry values of the gel degradation corresponding to the 92 kDa band was analyzed by ImageJ software, from the gels of 4 independent experiments. F. Azurophil granule exocytosis was determined by the increase in plasma membrane expression of the CD63 marker by flow cytometry. Data are expressed as the mean ± SEM of the mean channel of fluorescence (mcf) from 5 independent experiments. *p

Article Snippet: The release of MMP-9 (a marker for gelatinase granule exocytosis) was measured in the neutrophil supernatants, collected from the experimental conditions described above, by MMP-9 ELISA (Boster) following the manufacturer’s instructions; as well as the enzymatic activity analyzed by gelatin zymography assay as previously described ( ).

Techniques: Flow Cytometry, Cytometry, Expressing, Fluorescence, Enzyme-linked Immunosorbent Assay, Zymography, Activity Assay, Software, Marker

Journal: Oncogene

Article Title: Aberrant low expression of p85α in stromal fibroblasts promotes breast cancer cell metastasis through exosome-mediated paracrine Wnt10b

doi: 10.1038/onc.2017.100

Figure Lengend Snippet: P85α deletion converts fibroblasts into activated myofibroblasts exhibiting features of CAFs. ( a ) Western blot analysis of p85α, phospho-Akt, Akt and α-smooth muscle actin (α-SMA) expression in WT and p85α −/− fibroblasts. p-Akt, Phospho-Akt (Ser473); T-Akt, Akt (pan). ( b ) Immunofluorescence analysis of α-SMA expression in WT and p85α −/− fibroblasts. Scale bar, 50μm. ( c ) The area of the contracted gels induced by WT or p85α −/− fibroblasts. The appearance of the contracted gels that have four replications at 72 h is also shown. ( d ) The morphology of WT or p85α −/− fibroblasts in a dish or collagen lattice described in ( c ). ( e ) Immunofluorescence, western blot and fluorescence-activated cell sorting (FACS) analysis of CD44 expression in WT and p85α −/− fibroblasts. Scale bar, 20 μm. ( f ) Reverse transcription–PCR analysis of TGF-β family proteins and SDF-1. MMP2 and MMP9 expression in WT and p85α −/− fibroblasts. ( g ) Quantitative PCR analysis of TGF-β1, SDF-1, MMP2 and MMP9 expression in WT and p85α −/− fibroblasts. Data are the mean±s.e.m. ( h ) Enzyme-linked immunosorbent assay (ELISA) analysis of TGF-β1, SDF-1, MMP2 and MMP9 expression in conditioned medium of WT and p85α −/− fibroblasts. Data are the mean±s.e.m. ** P- value⩽0.01. ( i ) ELISA analysis of TGF-β1, SDF-1, MMP2 and MMP9 expression in p85α −/− fibroblast-conditioned medium of when treated with or without the PI3K inhibitor LY294002 (50 μ M ) for 24 h and the DMSO solution as a control. Con, p85α −/− fibroblasts treated with DMSO, LY, p85α −/− fibroblasts treated with LY294002. Data are the mean±s.e.m. ** P- value⩽0.01.

Article Snippet: The supernatants were measured using a commercially available SDF-1 ELISA Kit (R & D Systems, Minneapolis, MN, USA), a TGF-β1 ELISA Kit (Boster Bio-Tech, Wuhan, China), an MMP2 ELISA Kit (Boster Bio-Tech), an MMP9 ELISA Kit (Boster Bio-Tech) and a Wnt10b ELISA Kit (Cusabio Biotech, Wuhan, China).

Techniques: Western Blot, Expressing, Immunofluorescence, Fluorescence, FACS, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Oncology Letters

Article Title: Early changes in the apparent diffusion coefficient and MMP-9 expression of a cervical carcinoma U14 allograft model following irradiation

doi: 10.3892/ol.2017.7035

Figure Lengend Snippet: Correlation between mean ADC values and tumor volume, mean ADC values and MMP-9 expression. (A) No significant correlation was observed between mean ADC values and tumor volume (P > 0.05). (B) A significant correlation between mean ADC values and MMP-9 expression levels was observed (P=0.003; r=0.752). Values were calculated using a Spearman's rank correlation test. ADC, apparent diffusion coefficient; MMP-9, matrix metalloproteinase.

Article Snippet: Immunohistochemistry for MMP-9 expression.

Techniques: Expressing, Diffusion-based Assay

Journal: Oncology Letters

Article Title: Early changes in the apparent diffusion coefficient and MMP-9 expression of a cervical carcinoma U14 allograft model following irradiation

doi: 10.3892/ol.2017.7035

Figure Lengend Snippet: Histological changes and MMP-9 expression in irradiated tumors and untreated tumors. (A) Histologic examination of hematoxylin and eosin staining (magnification, 400×): i) Untreated tumor; ii) 6 h following irradiation; iii) 24 h following irradiation; iv) 72 h following irradiation. Irradiated tumors exhibited cellular edema, swelling, increases in size, cell layer loosening and intercellular space dilatation ii-iii). (B) MMP-9 expression determined via immunohistochemical staining (magnification, ×400). i) Untreated tumor; ii) 6 h following irradiation, MMP-9 expression in immunohistochemical staining was markedly increased in comparison with untreated tumor (IHC score: ++ vs. +); iii) 24 h following irradiation; iv) 72 h following irradiation. MMP-9, matrix metalloproteinase.

Article Snippet: Immunohistochemistry for MMP-9 expression.

Techniques: Expressing, Irradiation, Staining, Immunohistochemistry