human mmp 2 elisa kit  (Millipore)

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Targeting Loxosceles spider Sphingomyelinase D with small-molecule inhibitors as a potential therapeutic approach for loxoscelism

doi: 10.1080/14756366.2018.1546698

Figure Lengend Snippet: SMase D binding and cell death of human keratinocytes induced by Loxosceles venom. HaCaT cells (5 × 10 4 /well) were treated with 10 μg of L. laeta venom, preincubated or not with each inhibitor to evaluate cell death, binding of the SMase D to the keratinocyte cell membrane and secretion of MMP-2 and MMP-9. Cell death was evaluated after 72 h of incubation by the MTT method. The ability of the toxins to bind to the surface of keratinocytes was analysed by flow cytometry using a horse polyclonal serum against L. intermedia SMases D. Detection of MMP-2 and 9 was performed using Calbiochem MMP- 2 and 9 ELISA Kits. Data are represented as mean ± SEM of duplicates representative of three independent experiments. Statistically analysed by One-Way ANOVA followed by Tukey HSD test. (*)Significant difference in relation to the buffer ( p

Article Snippet: Supernatants of the keratinocyte cultures, treated for 72 h with venom in the presence or absence of inhibitors (5 μg), were centrifuged at 405 g for 10 min, and tested for the presence of MMP-2 and MMP-9, using the MMP-2 ELISA Kit (QIA63-1EA) and MMP-9 ELISA Kit (QIA56-1EA), according to the manufacturer’s instructions (Calbiochem®, USA).

Techniques: Binding Assay, Incubation, MTT Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Journal of Enzyme Inhibition and Medicinal Chemistry

Article Title: Targeting Loxosceles spider Sphingomyelinase D with small-molecule inhibitors as a potential therapeutic approach for loxoscelism

doi: 10.1080/14756366.2018.1546698

Figure Lengend Snippet: SMase D binding and cell death of human keratinocytes induced by Loxosceles venom. HaCaT cells (5 × 10 4 /well) were treated with 10 μg of L. laeta venom, preincubated or not with each inhibitor to evaluate cell death, binding of the SMase D to the keratinocyte cell membrane and secretion of MMP-2 and MMP-9. Cell death was evaluated after 72 h of incubation by the MTT method. The ability of the toxins to bind to the surface of keratinocytes was analysed by flow cytometry using a horse polyclonal serum against L. intermedia SMases D. Detection of MMP-2 and 9 was performed using Calbiochem MMP- 2 and 9 ELISA Kits. Data are represented as mean ± SEM of duplicates representative of three independent experiments. Statistically analysed by One-Way ANOVA followed by Tukey HSD test. (*)Significant difference in relation to the buffer ( p

Article Snippet: MMPs dosage in the culture supernatants from human keratinocytes Supernatants of the keratinocyte cultures, treated for 72 h with venom in the presence or absence of inhibitors (5 μg), were centrifuged at 405g for 10 min, and tested for the presence of MMP-2 and MMP-9, using the MMP-2 ELISA Kit (QIA63-1EA) and MMP-9 ELISA Kit (QIA56-1EA), according to the manufacturer’s instructions (Calbiochem®, USA).

Techniques: Binding Assay, Incubation, MTT Assay, Flow Cytometry, Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Science translational medicine

Article Title: Detection of early pancreatic ductal adenocarcinoma using thrombospondin-2 and CA19-9 blood markers

doi: 10.1126/scitranslmed.aah5583

Figure Lengend Snippet: Phase 1 validation studies and THBS2 expression in PDAC and other human tumors (A) AUC analysis of blinded ELISA data for the proteinsMMP2, MMP10, and THBS2 in plasma samples from 10 patients with PDAC at various stages of disease compared to 10 healthy controls. (B) Boxplots of THBS2 mRNA expression measured in various human tumors (sample sizes in parentheses) assessed by RNA-seq. Tumors are sorted in order of decreasing median expression of THBS2 mRNA. Of the pancreatic cancer samples from the TGCA database (n=179), we analyzed only PDAC (n=134). All expression values are log2(RSEM values =1) transformed.

Article Snippet: ELISA kits for human MMP2 (Millipore), human MMP10 (Ray Biotech), and human Thrombospondin-2 Quantikine (DTSP20, R & D systems) were used as described by the manufacturers’ instructions.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, RNA Sequencing Assay, Transformation Assay

Journal: Microorganisms

Article Title: Multidrug-Resistant Pseudomonas aeruginosa Evokes Differential Inflammatory Responses in Human Microglial and Retinal Pigment Epithelial Cells

doi: 10.3390/microorganisms8050735

Figure Lengend Snippet: CHME-3 and ARPE-19 cells were challenged with S- PA and MDR- PA , with a MOI of 10:1. At indicated time points, the cells were collected and processed for RNA isolation; cDNA synthesis followed by RT-qPCR. The bar graphs show that there was no significant expression of MMP-2 ( A , D ) or tissue inhibitor of metalloproteinases (TIMP)-1 ( B , E ) in either CHME-3 or ARPE-19 cells. ( C , F ) Significantly elevated expression of MMP-9 was observed after infection of CHME-3 and ARPE-19 cells with MDR- PA and S- PA strains. The data are shown as the mean ± SE from three sets of independent experiments; ** p

Article Snippet: The minimum detectable dose of Human MMP-2 was determined to be 3500 pg/mL, and MMP-9 was determined to be 10 pg/mL.

Techniques: Isolation, Quantitative RT-PCR, Expressing, Infection