Depolarization of Mmp (ΔΨm) is observed during early apoptosis (31 (link)). Changes in platelet Mmp were determined by staining with cationic dye 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1; Molecular Probes; Thermo Fisher Scientific, Inc). Specifically, 50 µl PRP was incubated with 500 µl JC-1 working solution at 37˚C with 5% CO2 for 10 min in the dark. Following filtration with a 70-µm cell strainer, samples were analyzed with a flow cytometer (BD Biosciences) Accuri C6) and FlowJo v.10.7. (BD Biosciences). A total of 20,000 events were acquired and platelets were detected in the platelet-specific gate. The results are presented as a ratio of the mean FL2 and FL1; decrease in the FL2:FL1 ratio (Mmp-Index) indicated a loss in Mmp (31 (link)). A parallel incubation in which carbonyl cyanide m-chlorophenylhydrazone (MedChemExpress) at 25 µM was added instead of JC-1 as control.
Carbonyl cyanide m chlorophenylhydrazone
Manufactured by MedChemExpress
Sourced in United States
Carbonyl cyanide m-chlorophenylhydrazone is a chemical compound used for research purposes. It is a cellular respiration uncoupler, which means it disrupts the process of ATP synthesis in mitochondria. This compound is commonly used in cell biology and biochemistry research.
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Lab products found in correlation
2 protocols using carbonyl cyanide m chlorophenylhydrazone
1
Measurement of Platelet Mitochondrial Membrane Potential
2
Antibiotic Susceptibility of Clinical Isolates
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Clinical isolates were obtained from a tertiary teaching hospital (Wenzhou, China). Reference strains (E. coli ATCC25922, K. pneumoniae ATCC700603, and A. baumannii ATCC19606) were used in all mechanism assays. Antibiotics including rifampicin, linezolid, erythromycin, tetracycline, vancomycin, and polymyxin B (PMB) (Kangtai, Zhejiang, China) were used in this study. Pentamidine, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and their solvent dimethyl sulfoxide (DMSO) were purchased from MedChemExpress (Junction, NJ, USA) and Sigma-Aldrich, USA (St. Louis, MO, USA). The broth microdilution (BMD) method was used to determine the MICs of all agents against all bacteria (Table S2 ). Sub-MICs of PMB (0.0625 mg/L) or SDS (0.01% v/v) (Solarbio, Beijing, China) were used as membrane permeators and CCCP (5 mg/L) as an efflux inhibitor [21 (link)]. Sub-MICs of CCCP (5 mg/L) and PMB (0.0625 mg/L) were added to serially two-fold-increased concentrations of linezolid, rifampicin, or vancomycin to determine their effects on the MICs of these antibiotics for multiple GNBs.
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