The lung tissue models were prepared as previously described (Braian et al., 2015 (link)). In brief, 16HBE cells and human primary macrophages (uninfected/infected at MOI 10 with Mtb expressing GFP), PKH26 red dye-labeled monocytes (stained according to manufacturer's instructions) were seeded into a matrix of collagen-embedded MRC-5 cells. During tissue development, the models were air-exposed at day 5 post infection allowing mucus secretion and stratification (Nguyen Hoang et al., 2012 (link); Parasa et al., 2014 (link)). At day 7 after infection, the models were harvested and fixed with 4% paraformaldehyde for 30 min. Supernatants were collected and stored at −80°C for cytokine and chemokine analysis. For MMP inhibition experiments, primary macrophages treated with a global MMP inhibitor, 200 nM marimastat (Merck Millipore), 16 h prior to Mtb infection were introduced in the lung tissue model. After the addition of macrophages, the tissue models were cultured in the presence of the inhibitor until tissue harvest.
Marimastat
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany
Marimastat is a laboratory equipment product developed by Merck Group. It is a synthetic matrix metalloproteinase (MMP) inhibitor designed for use in research and scientific applications. Marimastat functions by inhibiting the activity of various MMPs, which play a role in various biological processes. Further details on the intended use of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
Batimastat, by Merck Group (5 mentions)
Phorbol myristate acetate (pma), by Merck Group (4 mentions)
Ionomycin, by Merck Group (3 mentions)
Lactacystin, by Merck Group (3 mentions)
Chloroquine, by Merck Group (3 mentions)
Gi254023x, by Merck Group (3 mentions)
Hecd 1, by Abcam (2 mentions)
Gm6001, by Merck Group (2 mentions)
Polyclonal goat igg, by R&D Systems (2 mentions)
Blebbistatin, by Merck Group (2 mentions)
Foxp3 fix perm buffer set, by Thermo Fisher Scientific (2 mentions)
Ionomycin cell stimulation cocktail, by Thermo Fisher Scientific (2 mentions)
Nrp1 extracellular domain antibody, by R&D Systems (2 mentions)
Antibody to adam10, by Merck Group (2 mentions)
Uk370106, by Santa Cruz Biotechnology (2 mentions)
Trypan blue dye, by Merck Group (1 mentions)
Cytochalasin d, by Merck Group (1 mentions)
Y 27632, by Abcam (1 mentions)
Mmp 2 inhibitor 1, by Merck Group (1 mentions)
L 685 458, by Merck Group (1 mentions)
45 protocols using marimastat
1
Multifaceted Lung Tissue Model for Tuberculosis
2
Evaluating ECFC Viability with Marimastat
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ECFCs were isolated and grown as previously described [40 (link)]. Cell viability upon Marimastat (Merck) treatment was evaluated after 24 h by Trypan blue dye (Merck) exclusion assay. When Marimastat is tested, DMSO is always used as control. Human Retinal Endothelial Cells (HRECs) were gently provided by Dr. Lulli, Italy and grown in EGM-2 culture medium.
3
Branching Morphogenesis Pathway Inhibition
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To analyze the influence of specific pathways and processes during branching morphogenesis different inhibitors were used. To inhibit actin polymerization organoids were treated with 1 µg/ml Cytochalasin D (Merck). organoids were analyzed 4 h after treatment. To inhibit collagen degradation 10 µM Marimastat (Merck) was added to the organoid culture. According experiments were started directly after treatment. Inhibition of Rho kinase was achieved by using 10 µM Y-27632 (Abcam). Blocking of E-cadherin was performed by addition of an E-chadherin blocking-antibody HECD1 (Abcam) in a dilution of 1:50. Specifically, HECD1 was added continuously from day 5 to day 14.
4
Inhibition of MMP-2 Activity and Invasion
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Cells were plated in 6 well plates and treated with either 10 μM Marimastat (EMD Millipore, Billerica, MA), 1 μM MMP-2 Inhibitor I (Cis-9-octadecenoyl-N-hydroxylamide oleoyl-N-hydroxylamide, EMD Millipore), or the equivalent volume of DMSO for 24 hours. An invasion assay was performed as described above. Cells were plated in six-well tissue culture plates and transfected per manufacturer's instructions with 20 μM MMP-2 siRNA or control siRNA with 3 μL of Dharamafect2 (Thermo Scientific, formerly Dharmacon) for 48 hours
5
Intestinal Organoid Differentiation Protocol
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After the initial 48 h of mTeSR medium in the Organoplate, cells were exposed to definitive endoderm differentiation medium (DE) Table S1 . After additional 48 h the differentiation on the definitive endoderm was completed, the cultures were started on Hindgut differentiation medium on day 4 (HG) Table S1 . These conditions were maintained for 3 days. Cells were further directed towards the mature intestine with mature intestine medium (MI) Table S1 for an additional two to three weeks, with medium changes every 3 to 4 days (Table S1 ). An amount of 10 µM MMP-8 inhibtor (Fischer Scientific No. 44-423-7) or 10 µM Marimastat (MERCK,. M2699) was added to the cultures once the Hindgut stage of differentiation was started to reduce the invasion of cells into the ECM.
6
Inhibition of Matrix Degradation and Cell-Cell Adhesion
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Inhibitors were added at various developmental stages to the cell culture media. In particular, the media was enriched with the inhibitors directly prior to the experiment, which was then present throughout the whole measurement. Matrix degradation was inhibited by 10 μM Marimastat (Merck), and cell–cell adhesion was lowered by blocking E-Cadherin via HECD1 (Abcam) in a dilution of 1:25.
7
Cadherin-11 Modulation in Cellular Signaling
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The antibodies used are as follows: mouse IgG1 isotype control (MOPC-21, Bio X Cell); anti-cadherin-11 monoclonal antibody 23C6 and 3H10 [6 (link)]; anti-cadherin-11 monoclonal antibody 5B2H5 and polyclonal antibody #71-7600 (Life Technologies); anti-β-actin (AC-15, Sigma-Aldrich); anti-presenilin 1 (D39D1) and anti-presenilin 2 (Cell Signaling Technology, Danvers, MA, USA); anti-ADAM 10 (AB19026, EMD Millipore, Billerica, MA, USA). Cadherin-11-Fc and E-cadherin-Fc fusion proteins were generated as previously described [3 (link), 33 (link)]. The other reagents used are as follows: ionomycin (Sigma-Aldrich); human TNF-α and platelet-derived growth factor (PDGF)-BB (R&D Systems, Minneapolis, MN, USA); L-685,458, lactacystin, batimastat, (TAPI-2), marimastat, and GM6001 (EMD Millipore). Culture media was concentrated using iCON™ 9kD concentrators (Thermo Scientific, Grand Island, NY, USA).
8
FXYD5 Expression in Head and Neck Cancer
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Cell culture media with 10 µM Marimastat (Sigma-Aldrich), 10 µM MMP8 inhibitor (CAS 236403-25-1, Merch Millipore) or vehicle control (DMSO) were added to the cells for 24 h. Control and MMP8 + HSC-3 cells were detached with Versene buffer (0.48 mM EDTA in PBS) and gentle scraping. 200 000 cells per well were plated in a Nunc™ v-bottom 96-well plate (Thermo Fisher Scientific) in 1% bovine serum albumin (BSA) in PBS. The cells were fixed in 4% paraformaldehyde for 15 min on ice. After washing with 1% BSA/PBS three times, the cells were stained with 1 µg/ml polyclonal FXYD5 antibody (Acris Origene) in 1% BSA /PBS for 45 min on ice. 1× PBS and 1 µg/ml rabbit IgG (Dako, Glostrub, Germany) were used as controls. After washing as above, the cells were stained with allophycocyanin (APC)-conjugated anti-rabbit secondary antibody (Invitrogen, 1:200), washed as above and analysed with Accuri™ C6 Plus (BD Biosciences).
9
IL-6 Signaling Modulation in T Cell Activation
10
ECM Degradation by Collagenase I
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Degradation of ECM by collagenase type I (C2674 Sigma) was examined in presence of MMP inhibitors. ECM formed by 16000 HCF cultured for 7 days was used for degradation study. 100 μg of collagenase was incubated with TIMP3 or marimastat (Sigma‐Aldrich) in decellularized ECM in 96‐wells for 1 hour at 37°C with gentle shake. The wells were washed once with 100 μL of PBS, fixed with 4% paraformaldehyde for 15 minutes and immunostained for collagen type III.
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