C8138

Manufactured by Merck Group

Sourced in Switzerland

The C8138 is a laboratory instrument designed for the analysis and measurement of various samples. It is a versatile piece of equipment that can be used in a wide range of scientific and research applications. The core function of the C8138 is to provide accurate and reliable data for users, enabling them to make informed decisions in their work.

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Lab products found in correlation

C8273, by Merck Group (2 mentions) Ldh release, by Promega (1 mentions) Dansylcadaverine, by Merck Group (1 mentions) C4555, by Merck Group (1 mentions) Dynasore, by Merck Group (1 mentions) W1628, by Merck Group (1 mentions) D7693, by Merck Group (1 mentions) H3149, by Merck Group (1 mentions) Wortmannin, by Merck Group (1 mentions) N6261, by Merck Group (1 mentions) Via probe cell viability solution, by BD (1 mentions) Cytochalasin d, by Merck Group (1 mentions)

3 protocols using c8138

Chemicals and Infection Quantification Protocol

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Chemicals were purchased and stocks were made as follows:
Chlorpromazine: Sigma C8138 solubilized in water at 20 mg/ml
Tetrandrine: MedChemExpress HY-13764 solubilized in DMSO at 5 mM
EIPA: Sigma A3085 solubilized in DMSO at 20 mM
In all cases, the same amount of vehicle was used for different concentrations and controls. The different chemicals were added at the same time as virus addition. Infection was quantified by flow cytometry 24 hours after infections. Percent infection compared to control was calculated based on a minimum of three replicates.
Cytotoxicity assays were performed on cells treated in parallel with chemical or vehicle only. Cytotoxicity was determined by assessment of LDH release (Promega). For graphing purposes, percent viability was calculated by subtracting percent cytotoxicity from 100. All cytotoxicity assays were performed with a minimum of three replicates.

Gourronc F.A., Rebagliati M., Kramer-Riesberg B., Fleck A.M., Patten J.J., Geohegan-Barek K., Messingham K.N., Davey R.A., Maury W, & Klingelhutz A.J. (2022). Adipocytes are Susceptible to Ebola Virus Infection. Virology, 573, 12-22.

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Inhibitors of Endocytosis and Phagocytosis

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Different endocytotic inhibitors were tested for their optimal concentration, exposure time and cell impairment (Table 1, Figure 3). Inhibition of clathrin-mediated endocytosis in both cell types was tested with chlorpromazine hydrochloride (C8138, Sigma-Aldrich, Switzerland) and monoDansylcadaverine (Dansylcadaverine, D4008, Sigma-Aldrich). Inhibition of caveolin-mediated endocytosis was performed in both cell lines using 10 mM methyl-β-cyclodextrin (mβcd) (C4555, Sigma-Aldrich). A 4 µM cytochalasin D (C8273, Sigma-Aldrich) solution was used to inhibit phagocytosis and macropinocytosis in both cell lines.

Kuhn D.A., Vanhecke D., Michen B., Blank F., Gehr P., Petri-Fink A, & Rothen-Rutishauser B. (2014). Different endocytotic uptake mechanisms for nanoparticles in epithelial cells and macrophages. Beilstein Journal of Nanotechnology, 5, 1625-1636.

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Optimization of Extracellular Vesicle Uptake

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Cells were preincubated for 30 min at 37°C in complete medium containing Heparin (0.1–100 μg/ml; H3149; Sigma), Dynasore (20–160 μM; D7693; Sigma), Chlorpromazine (1–50 μM; C8138; Sigma), Nystatin (5–40 μg/ml; N6261; Sigma), Cytochalasin D (0·25–2 μM; C8273; Sigma), or Wortmannin (0.1–10 μM; W1628; Sigma). Cell viability was measured by flow cytometer using BD Via-Probe™ Cell Viability Solution containing 7-AAD. PKH-MP were then added to the cells (ratio 1:100,000) and analyzed after 6 h by flow cytometer. Following the results of these experiments, combination treatment with inhibitors of endocytosis (10 μg/ml Heparin and 80 μM Dynasore) and phagocytosis (5 μM Wortmannin) was also tested. Drug vehicle controls were used for the experiments: 0.1% PBS as a control for Heparin, Chlorpromazine, Nystatin, 0.1% dimethyl sulfoxide (DMSO) for Dynasore, Cytochalasin D, and Wortmannin.

da Costa Gonçalves F., Korevaar S.S., Ortiz Virumbrales M., Baan C.C., Reinders M.E., Merino A., Lombardo E, & Hoogduijn M.J. (2021). Mesenchymal Stromal Cell Derived Membrane Particles Are Internalized by Macrophages and Endothelial Cells Through Receptor-Mediated Endocytosis and Phagocytosis. Frontiers in Immunology, 12, 651109.

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