M7027

Manufactured by Merck Group

Sourced in United States

M7027 is a laboratory equipment product manufactured by Merck Group. It is designed for use in scientific research and analysis. The core function of M7027 is to provide a controlled environment for various laboratory processes and experiments.

Automatically generated - may contain errors

Lab products found in correlation

Cytochalasin d cyt d, by Merck Group (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) C1022, by Merck Group (1 mentions) A5040, by Merck Group (1 mentions) Fv3000, by Olympus (1 mentions) Cls3471, by Corning (1 mentions) Methylcellulose, by Merck Group (1 mentions) D 1841, by Thermo Fisher Scientific (1 mentions) Quemesa charge coupled device camera, by Olympus (1 mentions) Tetracycline, by Merck Group (1 mentions) Prism 8, by GraphPad (1 mentions) Pbs ph 7, by Thermo Fisher Scientific (1 mentions) Metformin hydrochloride, by Cayman Chemical (1 mentions)

10 protocols using m7027

Culturing BJ Cell Lines for Experimental Analyses

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The cell lines BJ (hTERT-immortalized human non-transformed fibroblast cells), BJ-LT (SV40 large T-antigen (LT)-transfected BJ cells), BJ-LT-Ras (BJ-LT cell transfected with oncogenic HRas mutant) [24 (link)] were cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, Life technologies, UK) supplemented with 10% fetal bovine serum (FBS, FB-1090–500, Werner Saveen, Biological Industries, Beit-Haemek Ltd, Israel), 100 U/ml penicillin and 0.1 mg/ml streptomycin (complete medium). The cells were kept at 37 °C in a humid atmosphere containing 5% CO₂. In some experiments 0.5–5 μM cytochalasin D (CytD, Sigma-Aldrich) were used to induce binucleation. Mitotic cells were collected by the shake off method in which exponentially growing cells were washed once with pre-warmed PBS, followed by incubation in complete medium for approximately 2–3 h, and then the loosely attached mitotic cells were detached by tapping the culture flasks. The cells were collected and re-suspended in fresh complete medium for culturing in either ultra-low attachment plates or on plates coated with fibronectin (40 μg/ml). Since BJ-LT cell had a higher tendency to aggregate via cell–cell contacts, 1.2% methylcellulose (M-7027, Sigma-Aldrich) was included in the culture medium when these cell lines were cultured in suspension in ultra-low attachment wells.

Gupta D.K., Kamranvar S.A., Du J., Liu L, & Johansson S. (2019). Septin and Ras regulate cytokinetic abscission in detached cells. Cell Division, 14, 8.

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Gastrointestinal Transit Time Assessment

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Male animals between 8–16 weeks were used, or as indicated in the manuscript when both female and male animals were examined. The night before the experiment, animals were transferred to individual housing with free access to water only. On the day of experiment, animals had free access to food and water for 1 hour. A solution of 6% carmine red (300 μl, Sigma, C1022) was prepared using 0.5% methylcellulose (Sigma, M7027) and was administered by gavage through a 21-gauge round-tip feeding needle (Roboz Surgical Instrument, FN-7903). 90 min after gavage, fecal pellets were monitored for the presence of carmine red. Total GI transit time was calculated from the time of administration to the first observance of carmine red in stool.

Song Y., Fothergill L.J., Lee K.S., Liu B.Y., Koo A., Perelis M., Diwakarla S., Callaghan B., Huang J., Wykosky J., Furness J.B, & Yeo G.W. (2023). Stratification of enterochromaffin cells by single-cell expression analysis. bioRxiv.

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Zebrafish Neuronal and Vascular Imaging

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For imaging, live zebrafish larvae anesthetized with 200 mg/L tricaine (Sigma-Aldrich, A5040, United States) were mounted in 3% methylcellulose (Sigma-Aldrich, M7027, United States). Brains were dissected under a dissection microscope (Olympus, DP73, Japan) at the time point of 4 dpf. Fixed samples were balanced and located in 80% glycerol before imaging. Images of neurons in the diencephalon area of Tg (Huc: RFP) and Tg (serpini1^−/−- HuC: RFP^+/+), and vascular of Tg (Fli1: EGFP) and Tg (serpini1^−/−- Fli1: EGFP^+/+) were acquired from a confocal microscope system (Olympus, FV3000, Japan). Using the Z stack strategy, top and bottom of the structure of interest were verified and a 2 um interval was selected to obtain the high-resolution images. Image procession and intensity measurements were done using ImageJ software (n = 10 for each group).

Han S., Zhang D., Dong Q., Wang X, & Wang L. (2021). Deficiency in Neuroserpin Exacerbates CoCl2 Induced Hypoxic Injury in the Zebrafish Model by Increased Oxidative Stress. Frontiers in Pharmacology, 12, 632662.

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3D Spheroid Assay for CRC Cells

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Anchorage-independent growth and spheroid formation of CRC cells was investigated in a 3D spheroid model. Briefly, cells (1000–4000 viable cells/well) were seeded into 6-well plates with ultra-low attachment surface (No. CLS3471, Corning Inc., Corning, NY, USA) in 2 mL of serum-free medium supplemented with epidermal growth factor (EGF) (100 ng/μL, No. AF-100-15), vascular endothelial growth factor (VEGF) (10 ng/μL, No. AF-100-20A) (PeproTech, Cranbury, NJ, USA) and 0.5% methylcellulose (No. M7027, Sigma–Aldrich). The cell spheres were allowed to grow for 10–15 days and imaged under an inverted fluorescence microscope. The number of spheres was counted and sphere size was assessed using ImageJ.

Yue X., Liu T., Wang X., Wu W., Wen G., Yi Y., Wu J., Wang Z., Zhan W., Wu R., Meng Y., Cao Z., Le L., Qiu W., Zhang X., Li Z., Chen Y., Wan G., Bu X., Peng Z, & Liu R.Y. (2023). Pharmacological inhibition of BAP1 recruits HERC2 to competitively dissociate BRCA1–BARD1, suppresses DNA repair and sensitizes CRC to radiotherapy. Acta Pharmaceutica Sinica. B, 13(8), 3382-3399.

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Measuring Gastric Emptying and Intestinal Transit

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Gastric emptying and small intestinal transit were measured by
evaluating the intestinal location of Rhodamine B-conjugated dextran (D1841,
Molecular Probes) as described elsewhere^{32 (link)} with modification. Six-week-old mice were given
2.5mg/ml of Rhodamine B-conjugated dextran in 5% methylcellulose (M7027, Sigma)
via gavage. Ten minutes after administration for the gastric emptying assay and
60 minutes after administration for the small intestinal transit assay, the
entire small bowel (unflushed) was divided into 10 equal segments and placed
into tubes with 4ml of PBS. The stomach was prepared similarly. Tissues were
homogenized and were centrifuged at 2,000 rpm for ten minutes to separate out a
pellet and supernatant. The fluorescence in each aliquot of the cleared
supernatant was read by a Synergy4 plate reader (excitation 540 nm/emission 625
nm, BioTek). Gastric emptying was calculated as the ratio of the total
fluorescent intensity in small intestine divided by total fluorescent intensity
in stomach and small intestine. For small intestinal transit, fluorescent
intensity for each small intestinal segment (I₁ to
I_10, numbered from oral side) was used to
calculate the geometric center of delivered dextran by the formula
below^{32 (link)}.

g e o m e t r i c c e n t e r = \sum_{n = 1}^{10} \frac{I_{n} \times n}{I_{1} + I_{2} + \dots + I_{10}}

Kondo J., Powell A.E., Wang Y., Musser M.A., Southard-Smith E.M., Franklin J.L, & Coffey R.J. (2015). LRIG1 Regulates Ontogeny of Smooth Muscle-Derived Subsets of Interstitial Cells of Cajal in Mice. Gastroenterology, 149(2), 407-419.e8.

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GEM Treatment in Murine Infection

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GEM was dissolved in dimethyl sulfoxide (DMSO) followed by dilution in 0.1% methyl cellulose (Sigma-Aldrich, M7027) and was used at 15 mg/kg body weight. Mice were treated with vehicle (0.1% methyl cellulose) or GEM once daily via gavage from days 3 to 7 after infection.

Kim Y.S., Kim J.K., Hanh B.T., Kim S.Y., Kim H.J., Kim Y.J., Jeon S.M., Park C.R., Oh G.T., Park J.W., Kim J.M., Jang J, & Jo E.K. (2020). The Peroxisome Proliferator-Activated Receptor α- Agonist Gemfibrozil Promotes Defense Against Mycobacterium abscessus Infections. Cells, 9(3), 648.

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Transmission Electron Microscopy for EV Analysis

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EV preparations obtained from blood plasma and urine were qualitatively analyzed with transmission electron microscopy (TEM) (Fig. 1E). Samples were deposited on a formvar coated grids stabilized with evaporated carbon film and glow discharged before sample application (AGS162-3 H, Agar Scientific). Neutral uranyl acetate (2% in AD) (21447-25, Polysciences) was used for staining after which grids were coated with 2% methyl cellulose (M7027, Sigma-Aldrich) / uranyl acetate (0,4%) solution. These grids were examined using a Tecnai G2 Spirit transmission electron microscope (Thermo Fisher Scientific FEI) operated at 100 kV and images were captured with a Quemesa charge-coupled device camera (Olympus Soft Imaging Solutions).
Information on density measurement, interface mixing, and characterization methods (Fig. 1E) of rEV (fNTA and anti-p24 ELISA) and EV (mass-spectrometry based proteomics and TEM) is provided in the Additional file 1.

Van Dorpe S., Lippens L., Boiy R., Pinheiro C., Vergauwen G., Rappu P., Miinalainen I., Tummers P., Denys H., De Wever O, & Hendrix A. (2023). Integrating automated liquid handling in the separation workflow of extracellular vesicles enhances specificity and reproducibility. Journal of Nanobiotechnology, 21, 157.

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Zebrafish Infection Model for Staphylococcus aureus

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Staphylococcus aureus SH1000 expressing mCherry (SH1000 pMV158-mCherry) [28 (link)] was cultured in brain heart infusion (BHI) broth medium (Sigma, 53286) at 37°C supplemented with tetracycline (Sigma, 87128) at 5 µg/ml. Zebrafish larvae at 30 hpf (hours post-fertilization) were microinjected into the circulation with bacteria as previously described [25 (link)]. Briefly, anesthetized larvae were embedded in 3% w/v methylcellulose (Sigma, M7027) and injected individually using microcapillary pipettes (WPI, TW100-4) filled with the bacterial suspension of known concentration. For macrophage depletion, clodronate liposomes (Liposoma BV) were injected at 26 hpf, as previously described [38 ]. Following infection, larvae were observed frequently up to 120 hpf, and numbers of dead embryos recorded at each time point.

Prajsnar T.K., Serba J.J., Dekker B.M., Gibson J.F., Masud S., Fleming A., Johnston S.A., Renshaw S.A, & Meijer A.H. (2020). The autophagic response to Staphylococcus aureus provides an intracellular niche in neutrophils. Autophagy, 17(4), 888-902.

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Optomotor Response in Zebrafish

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Nine dpf zebrafish were embedded in 3% methylcellulose (Sigma Aldrich M7027) in a WillCo-dish® GWSt-3522. The WillCo-dish was placed inside the platform (OKR apparatus) which contains a drum (10 cm in diameter) lined with a white paper that has five vertical black stripes (1 cm in width), the drum rotated clockwise (1 stimulus per second), and eye movement was observed using a stereomicroscope positioned above the device. Recordings of 30 seconds per zebrafish and the number of movements of both the left and right eye per zebrafish were counted and analysed by two independent individuals (Brockerhoff 2006) (link). A blank white paper without black stripes was used as a control for the zebrafish response. Results were analysed for statistical significance using GraphPad Prism 8.4.3 software (GraphPad, San Diego, CA, USA). The mutant group (tapt1a -/-;tapt1b -/-) was obtained based on phenotyping the zebrafish by looking at the eye phenotype and pigmentation pattern (n = 20), while the control group in this experiment was represented by the rest of the clutch (n = 20), which were tapt1a +/+ ;tapt1b +/+ , tapt1a -/+ ;tapt1b -/+ , tapt1a -/+ ;tapt1b +/+ , tapt1a +/+ ;tapt1b -/+ , tapt1a -/-;tapt1b +/+ , tapt1a +/+ ;tapt1b -/-. Mutant tapt1a -/-;tapt1b -/-genotypes were confirmed by Sanger sequencing after the experiment.

Jarayseh T., Guillemyn B., De Saffel H., Bek J.W., Syx D., Symoens S., Gansemans Y., Van Nieuwerburgh F., Jagadeesh S., Raja J., Malfait F., Coucke P.J., De Clercq A, & Willaert A. (2023). A tapt1 knock-out zebrafish line with aberrant lens development and impaired vision models human early-onset cataract. Human genetics, 142(3).

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Formulation and Dosing of Metformin and O304

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The drug vehicle of 2% MC (M7027; Sigma-Aldrich, St. Louis, Missouri, USA) was formulated by adding 1.2 g MC powder to 60 mL cold phosphate-buffered saline (PBS) pH 7.4 (Fisher Scientific, Pittsburgh, Pennsylvania, USA) and stirring vigorously on a magnetic stirrer until clear. Metformin was formulated by adding 240 mg of metformin hydrochloride (Cayman Chemical, Ann Arbor, Michigan, USA) to 60 mL cold MC solution and stirring vigorously on a magnetic stirrer until clear, to a final concentration of 4 mg/mL. O304 was formulated by adding 240 mg of O304 (gift from Baltic Bio AB, Umeå, Sweden) to 60 mL cold MC solution and stirring vigorously on a magnetic stirrer for 60 min to produce a white suspension at a final concentration of 4 mg/mL. The combined drugs were formulated by adding 240 mg of metformin hydrochloride and 240 mg of O304 to 60 mL cold MC solution and stirring vigorously on a magnetic stirrer for 60 min to produce a white suspension at a final concentration of 4 mg/mL for each drug. A daily oral gavage of 1.0 mL delivers 4 mg of O304 and/or metformin, which is 200 mg/kg in a 0.02 kg mouse.

Das V., Kroin J.S., Moric M., McCarthy R.J, & Buvanendran A. (2019). Antihyperalgesia effect of AMP-activated protein kinase (AMPK) activators in a mouse model of postoperative pain. Regional anesthesia and pain medicine.

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