The curcumin derivatives, including CB-2, were synthesized in our laboratory. The high-performance liquid chromatography (HPLC) purity of these compounds was at least 99%. Dimethyl sulfoxide (DMSO) was used to prepare the 0.1-M CB-2 stock solution, followed by preservation at −20 °C and dilution to diverse concentrations using a suitable medium prior to utilization. Fetal bovine serum (FBS) was acquired from Hyclone Laboratories (Logan, UT, USA). Both DMEM and RPMI-1640 medium were provided by Gibco, Life Technologies (Carlsbad, CA, USA). Wortmannin (Wort, S1952), DCFH-DA (S0033), N-acetylcysteine (NAC; S0077), and JC-1 (C2006) were provided by Beyotime Biotechnology (Shanghai, China). An Annexin V-FITC/PI apoptosis detection kit (556547) was acquired from BD-Pharmingen (Franklin Lakes, NJ, USA). Earle’s Balanced Salt Solution (EBSS; E2888), thiazolyl blue tetrazolium bromide (MTT; M2128), Protease Inhibitor Cocktail (P8340), and diphenyleneiodonium chloride (DPI; D2926) were provided by Sigma-Aldrich (St. Louis, MO, USA). Paraformaldehyde (E672002-0500) was obtained from Shanghai Sangon Biotech (Shanghai, China). The lactate dehydrogenase detection kit was purchased from the JianCheng Bioengineering Institute (Nanjing, China).
Jc 1 c2006
Manufactured by Beyotime
Sourced in China
The JC-1 (C2006) is a fluorescent dye used for the detection of mitochondrial membrane potential in cells. It can be used to monitor changes in mitochondrial membrane potential, which is an indicator of cellular health and function.
Automatically generated - may contain errors
Lab products found in correlation
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Cytiva (1 mentions)
Paraformaldehyde (pfa), by Sangon (1 mentions)
Annexin 5 fitc pi apoptosis detection kit, by BD (1 mentions)
Ix71 fluorescence microscope, by Olympus (1 mentions)
Ferrostatin 1 fer 1, by Selleck Chemicals (1 mentions)
Gb12001, by Wuhan Servicebio Technology (1 mentions)
Gb23303, by Wuhan Servicebio Technology (1 mentions)
Gb23301, by Wuhan Servicebio Technology (1 mentions)
Phenylephrine pe, by Merck Group (1 mentions)
A12128, by ABclonal (1 mentions)
A1933, by ABclonal (1 mentions)
E bc k773 m, by Elabscience (1 mentions)
A1002, by ABclonal (1 mentions)
A006 2 1, by Nanjing Jiancheng (1 mentions)
A003 4 1, by Nanjing Jiancheng (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Phospho c raf ser259, by Cell Signaling Technology (1 mentions)
Cyclin d1, by Cell Signaling Technology (1 mentions)
7 protocols using jc 1 c2006
1
Curcumin Derivatives Synthesis and Characterization
2
Mitochondrial Membrane Potential Assay
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The mitochondrial membrane potential assay kit (JC-1, C2006; Beyotime) was used to measure the mitochondrial membrane potential (MMP) of the HepG2 cells according to the manufacturer’s instructions. Briefly, the cells were washed twice with PBS and then incubated with JC-1 at 37°C for 30 min. After incubation, the cells were washed again. An IX71 fluorescence microscope (Olympus) was used to observe the difference between green and red fluorescence. Red and green fluorescence represent high and low MMP, respectively.
3
Mitochondrial membrane potential assay
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Staining with a mitochondrial membrane potential assay kit (JC-1 C2006, Beyotime) was performed at 37°C for 20 min to determine the BMSC mitochondrial membrane potential.
4
PQQ Mitigates Oxidative Stress
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PQQ was obtained in the form of a disodium salt from Nascent Health Sciences LCC (NY, USA). Phenylephrine (PE) was from Sigma-Aldrich (MO, USA). Primary antibodies specific for YAP, GPX4, FSP1, and CoQ10 were from ABclonal (A1002, A1933, A12128, and A15193), while antibodies specific for p-YAP, BNP, β-actin, GAPDH, and Histone H3, as well as secondary goat anti-rabbit and goat anti-mouse IgG were from Wuhan Servicebio Technology Co., Ltd. (GB114060, GB11667, GB12001, GB15002, GB11026, GB23303, and GB23301). The 2′,7′-dichlorofluorescein diacetate (DCFH-DA) (S0033) and the mitochondrial membrane potential assay kit JC-1 (C2006) were purchased from the Beyotime Institute of Biotechnology. A test kit for measuring ferrous ion concentration (E-BC-K773-M) was purchased from Elabscience (Wuhan, China), and the kits for measuring malondialdehyde (MDA) and glutathione (GSH) levels were obtained from Nanjing Jiancheng Institute of Biological Engineering (A003-4-1, A006-2–1). Ferrostatin-1(Fer-1) was purchased from Selleck Chemicals (USA). Gibco (CA, United States) was the source of all cell culture reagents.
5
Celastrol's Impact on Mitochondrial Potential
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Cultured NRVMs were exposed to various concentrations of celastrol for 24 h. Then the mitochondrial membrane potential (ΔΨm) was measured using a mitochondrial membrane potential assay kit (JC-1, C2006; Beyotime Biotechnology) according to the manufacturer’s protocol.
6
Rhein Induces Apoptosis via PI3K/AKT Pathway
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The following antibodies and reagents were purchased: CDK4 (#12790), CDK6 (#13331), cyclin D1 (#55506), cyclin E (#20808), p21 (#2947), p27 (#3686), PARP (#9532), caspase-9 (#9504), cleaved-caspase-3 (#9664), AKT (#4691), phospho-AKT (Ser473) (#4060), phospho-PDK1 (Ser241) (#3438), PTEN (#9188), phospho-PTEN (Ser380) (#9551) and phospho-c-raf (Ser259) (#9421) antibodies were purchased from Cell Signaling Technology (Beverly, MA, USA). Bcl-2 (ab32124), Bcl-XL (ab32370), Bax (ab32503) and cytochrome c (ab133504) antibodies were purchased from abcam. GAPDH (60004-1-Ig), Survivin (10508-1-AP), XIAP (66800-1-Ig), COXIV (11242-1-AP) and PCNA (60097-1-Ig) antibodies were purchased from Proteintech Group (Chicago, IL, USA). Rhein (R7269), Oxaliplatin (O9512) and N-acetyl-L-cysteine (NAC, A7250) were obtained from Sigma-Aldrich (St. Louis, MO, USA). DCFH-DA (S0033) and JC-1 (C2006) were purchased from Beyotime (Haimen, China). Z-DEVD-FMK (S7312), z-LEHD-FMK (S7313) and z-VAD-FMK (S8102) were purchased from Selleck (Houston, TX, USA). LY294002 (HY-10108) and 740Y-P (HY-P0175) were purchased from MCE (Monmouth, NJ, USA).
7
Assessing Mitochondrial Membrane Potential
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Mitochondrial membrane potential detection reagent kit (JC-1; C2006) was purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). The cells were incubated with 5% CO2 at 37 °C for 3 h, added with 500 μL JC-1 working fluid (500 μL 1 × Incubation Buffer + l μL JC-1) in 5% CO2 at 37 °C for 15–20 min, rinsed with 1 × Incubation Buffer, and observed under a fluorescence microscope (XSP-63B, Shanghai Optical Instrument Factory, Shanghai, China). The results were scored as follows: normal cells: observation of double color filter: green ++red ++ (high green high red), the color under the same color filter was yellow green; apoptotic cells: observation of double color filter: green ++ red + (high green low red), the color under the same color filter was green.
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