Invasion assays were performed using a BD Matrigel® Invasion Chamber 24-well plate (BD Biosciences, San Jose, CA, USA). T-REx HeLa stable cell lines pre-cultured in medium containing 1 µg/ml doxycycline (DOX) for 24 h were seeded at a density of ~ 1 × 105 cells/well in the upper chamber of the transwell with SFM containing 1 µg/ml DOX. The lower chamber was filled with a standard culture medium containing 1 µg/ml DOX. Invading cells were stained with Diff-Quik reagents (Sysmex, Kobe, Japan) and then counted. An MMP-12 inhibitor, MMP408, was purchased from Merck KGaA (Darmstadt, Germany).
Mmp408
Manufactured by Merck Group
Sourced in Germany
The MMP408 is a versatile lab equipment designed for various research and testing applications. It functions as a mechanical microtiter plate mixer, capable of precisely mixing and agitating samples in microplates. The device ensures consistent and reliable sample preparation, making it a valuable tool for researchers and scientists across various disciplines.
Automatically generated - may contain errors
Lab products found in correlation
Diff quik reagent, by Sysmex (1 mentions)
Avance 3 700 mhz, by Bruker (1 mentions)
Combiflash, by Teledyne (1 mentions)
Gm6001, by Enzo Life Sciences (1 mentions)
Chromnav system, by Jasco (1 mentions)
Luna c18 10μ 10 250mm, by Phenomenex (1 mentions)
Lps from e coli o111 b4, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Latrunculin a, by Merck Group (1 mentions)
α tubulin t5168, by Merck Group (1 mentions)
Tgf β, by Enzo Life Sciences (1 mentions)
12 protocols using mmp408
1
Matrigel Invasion Assay for HeLa Cells
2
Analytical Characterization of Small Molecules
3
Modulating MMP-12 in HDM-induced RSV Infection
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Four days after the last HDM injection in the HDM group, mice were injected with 100 μg of the selective MMP-12 inhibitor MMP408 (Merck, Darmstadt, Germany) or hydroxypropyl methylcellulose (HPMC) via gastrointestinal (g.i.) injections. Two hours later, mice were infected with RSV (1 × 106 PFU) via i.n. injections. Mice were treated with MMP408 or HPMC via g.i. injections every 24 h after RSV infection. All mice were assessed on day 4 after RSV infection. In a separate experiment, 5 days after the last HDM injection in the HDM group (2 days after RSV infection), mice were treated with rMMP-12 (1 μg/30 μl; R&D systems) or PBS via i.n. injections every 24 h. All mice were assessed on day 4 after RSV infection.
4
Inhibiting MMP-12 in Colorectal Cancer
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MMP408 (MMP12 inhibitor, Cat.: 444291, Merck), CAS 1258003-93-8, could control the biological activity of MMP-12 [33 (link), 34 (link)]. All 17-week-old ApcMin/+ mice were randomly divided into three groups (n = 5 per group). One group of ApcMin/+ mice were intragastrically administered with MMP408 at a dose of 5 mg/kg, and the other group was intraperitoneally administered with 5-FU (30 mg/kg) combined with a dose of MMP408 by intragastric administration. Meanwhile, ApcMin/+ mice injected with normal saline as the control. Body weight was measured after the administration every 2 days, which continued for 10 days. Weight changes were recorded.
5
Modulation of Serum Endoglin Levels
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Ten-week-old mice were injected (i.p.) with 1 mg/kg body weight of lipopolysaccharide (LPS; from E. coli O111:B:4, Sigma-Aldrich, St. Louis, MO, USA) or PBS as a control condition. Three hours before LPS injection, mice were orally gavaged with 150–200 µL of 0.5% methylcellulose, 2% Tween-80 (vehicle), or MMP-408 (30 mg/kg body weight; Merck-Millipore, Burlington, MA, USA). Vehicle or MMP-408 gavage was repeated 24 h after i.p. injection with LPS. All animals were handled in strict accordance with good animal practice as defined by the national animal welfare bodies (RD 1201/2005 BOE #252). The experimental design and all animal work were approved by our institutional Ethical Committee for animal experiments. Mice were deeply anesthetized, and blood samples were obtained by puncture of posterior vena cava. The blood was centrifuged for 15 min at 1000g to collect serum samples. Levels of mouse sEng in serum were determined by DuoSet® ELISA Mouse Endoglin/CD105 Immunoassay (R&D Systems, Minneapolis, MN, USA).
6
Rat Cardiac Cell Culture Protocols
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All cell culture reagents for the growth of rat cardiac cells were purchased from Invitrogen, unless otherwise stated. Cell culture plastic was obtained from Greiner, Sarstedt or Nunc. All fine chemicals were of highest quality available and were purchased from Sigma Aldrich (DAPI, FITC-phalloidin, Hoechst 33342, β-aminopropionitrile, CCG-203971), Biomol (Fasudil, H1152P, SR-3677), Preprotech (TGF-β), Enzo Life Sciences (Latrunculin A) or Merck (MMP 408). Antibodies were purchased from BD Biosciences (ROCK1, 611,136), Santa Cruz (ROCK2, sc-5561), Origene (LOX, TA337077), Zytomed Systems (GAPDH, RGM2-6C5) and Sigma Aldrich (α-tubulin, T 5168; β-actin, A 2228; α-smooth muscle Actin (SMA), A 5228). All primers were synthesized by Eurofins Genomics.
7
Investigating MMP12 Role in Subretinal Fibrosis
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To investigate the role of MMP12 in subretinal fibrosis, an MMP12 specific inhibitor MMP408 [20 (link)] was used in the two-stage laser-induced mouse model of subretinal fibrosis. Three days after the second laser, mice were randomized into 3 groups, MMP408 treatment group, vehicle (1% dimethyl sulfoxide (DMSO) and 2% Tween 80) treatment group, and control non-treatment group. 5 mice were used in each group. MMP408 (Cat: 444291, Sigma Aldrich) was administered via gavage feeding at 5 mg/kg (150 µl/mouse) twice daily for five consecutive days. Mice in the vehicle group received the same amount of 1% DMSO and 2% Tween 80 for five days. Mice were sacrificed 10 days after the second laser and eyes were collected and processed for immunohistochemistry investigations.
8
Evaluating MMP408's Impact on ACP Cell Proliferation
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ACP cells were seeded into 24-well plates at a density of 3×103 cells/well, and in 9 wells, cells were treated with 2 nmol/ml MMP408 (37°C, Sigma-Aldrich; Merck KGaA) for 24, 48 or 72 h (three wells/condition) as the treatment group, and the remaining wells were treated for different periods of time (24, 48 or 72 h) at 37°C, as the control group. DMEM solution containing 10 µl Cell Counting Kit-8 (CCK-8, 10:1, cat. no. 40203ES60; Shanghai Yeasen Biotechnology Co., Ltd.) was added to each well and cells were incubated for an additional 2.5 h in the dark. Cell proliferation was subsequently analyzed at a wavelength of 450 nm, using a microplate reader (Enspire 23001489, PerkinElmer, Inc.). The number of cells in each treatment condition was determined, based on the absorbance.
9
Wound Healing Assay for Cell Migration
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The wound healing assay was performed to assess the migratory ability of ACP cells. ACP cells were seeded into 6-well plates at a density of 5×105 cells/well and incubated in media supplemented with 15% FBS (37 (link)) (Gibco; Thermo Fisher Scientific, Inc.), at 37°C for 4 h. Once the cells reached confluence (the cells cover the plate), the monolayers were scratched with a 200 µl pipette tip and washed three times with PBS. Cells were incubated in DMEM supplemented with 15% FBS (37 (link)) at 37°C for 48 h (Gibco; Thermo Fisher Scientific, Inc.) and the treatment group was treated with 2 nmol/ml MMP408 (Sigma-Aldrich; Merck KGaA) at 37°C for 48 g. Images were taken at 0 and 48 h using an inverted fluorescence microscope (magnification, ×200).
10
MMP408 Inhibits ACP Cell Growth
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ACP cells were seeded into6-wells plates at a density of 500 cells/well, with three wells as the control group and the other three wells as the treatment group. All cells were incubated in DMEM supplemented with 15% FBS (37 (link)) (Gibco; Thermo Fisher Scientific, Inc.). at 37°C for 24 h. The treatment groups were treated with 2 nmol/ml MMP408 at 37°C for 2 weeks (Sigma-Aldrich; Merck KGaA). Cells were cultured for 2 weeks, fixed with 4% paraformaldehyde for 30 min at 23°C stained with 50% crystal violet dye solution at 23°C for 2 h and imaged. Cell colonies were observed under a fluorescence microscope (magnification, ×200); a colony was counted if it contained ≥50 cells.
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