Compounds 1–6 (purity > 95%, Figure 1 ) were isolated from S. sonchifolius leaf as described in a previous report by Kitai et al [12 (link)]. Etoposide, taxol, and parthenolide were purchased from Sigma-Aldrich Co. Ltd., (Tokyo, Japan). Propidium iodide (PI), RNase A, and 4′, 6-diamidino-2-phenylindole (DAPI) were obtained from Sigma-Aldrich Co. Ltd., (Tokyo, Japan). A JC-1 assay kit was purchased from Cayman Chemical Co. Ltd., (Ann Arbor, MI, USA). Anti-cytochrome c and immunoglobulin G horseradish peroxidase (HRP) antibody were obtained from Santa Cruz Biotechnology, Inc., (Santa Cruz, CA, USA). A β-actin antibody was purchased from Gene Tex, Inc., (Irvine, CA, USA).
Jc 1 assay kit
Manufactured by Cayman Chemical
Sourced in United States
The JC-1 assay kit is a fluorescent probe used to detect changes in the mitochondrial membrane potential (ΔΨm) of cells. The kit measures the ratio of J-aggregates to monomeric JC-1, which is an indicator of ΔΨm. This assay can be used to study various cellular processes related to mitochondrial function.
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2 protocols using jc 1 assay kit
1
Isolation and Evaluation of Antiproliferative Compounds
2
Measuring Mitochondrial Membrane Potential
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As an early event in the mitochondrial apoptosis cascade, mitochondrial membrane potential (MMP) depolarization was measured using JC‐1 assay kit (Cayman, USA). Briefly, after experimental treatment, AML12 cells were cultured in 12‐well plates and incubated with JC‐1 staining solution (5 μg/ml) at 37°C for 15 min and washed twice with PBS. MMP was estimated by measuring the fluorescence ratio of free JC‐1 monomers (green) to JC‐1 aggregates in mitochondria (red) using fluorescence microscopy (Leica, DMLB2, Germany). Mitochondrial depolarization is indicated by an elevation in the ratio of hepatocyte emitting green fluorescence.
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