Calcein-AM, curcumin, deferoxamine, 8-hydroxyquinoline (8-HQ), ferric ammonium citrate (FAC), propidium iodide (PI), N-acetylcysteine (NAC) were from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Calcein-AM, 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), 3,3′- dihexyloxacarbocyanine iodide [DiOC6(3)] and N-[4-[6-[(acetyloxy) methoxy]-2,7-dichloro-3-oxo-3H-xanthen-9-yl]-2-[2-[2-[bis[2[(acetyloxy) methoxy]-2-oxyethyl] amino]-5-methyl-phenoxy] ethoxy]phenyl- N-[2-[(acetyloxy) methoxy]-2-oxyethyl]-(acetyloxy) methyl ester (Fluo-4/AM) were from Molecular Probes (Invitrogen, Eugene, OR, USA). Agarose was from Lonza (Walkersville, MD, USA. 8-Hydroxyquinoline was given together with FAC to enhance its internalization.
3 3 dihexyloxacarbocyanine iodide
Manufactured by Thermo Fisher Scientific
Sourced in United States
3,3′-Dihexyloxacarbocyanine Iodide is a fluorescent dye used for labeling and tracking cells in various biological applications. It is a lipophilic cation that can passively diffuse across cell membranes and accumulate in the mitochondria of living cells. The dye exhibits green fluorescence when excited by blue or violet light.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Propidium iodide, by Merck Group (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Deferoxamine, by Merck Group (1 mentions)
2 7 dichlorodihydrofluorescein diacetate dcfh da, by Thermo Fisher Scientific (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
Calcein am, by Merck Group (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Curcumin, by Merck Group (1 mentions)
Agarose, by Lonza (1 mentions)
Facscalibur, by Merck Group (1 mentions)
Human alpha thrombin, by Merck Group (1 mentions)
Lsm 880, by Zeiss (1 mentions)
Calpeptin, by Merck Group (1 mentions)
Trypan blue, by Merck Group (1 mentions)
Mitochondria cytosol fractionation kit, by Abcam (1 mentions)
Caspase 3 and caspase 9 colorimetric assay kits, by R&D Systems (1 mentions)
1 2 bis 2 aminophenoxy ethane n n n n tetraacetic acid tetrakis acetoxymethyl ester bapta am, by Merck Group (1 mentions)
5 protocols using 3 3 dihexyloxacarbocyanine iodide
1
Assessing Cellular Oxidative Stress
2
Apoptosis and Mitochondrial Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HCC cells were treated with DMSO or armillarikin, collected, and then resuspended in hypotonic buffer (0.1% each of sodium citrate and Triton X-100) containing 5 µg/mL propidium iodide (Sigma) as described by Nicoletti et al19 (link) to detect the apoptotic cells with hypodiploid chromosomal DNA (sub-G1 or DNA laddering) by using flow cytometry (fluorescence-activated cell sorting, FACSCalibur).20 The percentage of the sub-G1 cells was calculated. To detect the collapsed mitochondrial transmembrane potential of the cells by using flow cytometry, they were mixed with 0.02 µM 3,3′-dihexyloxacarbocyanine iodide (Molecular Probes, Eugene, OR, USA) and analyzed according to the manufacturer’s instructions. All the flow cytometric data were analyzed by using the FlowJo software (Tree Star, Inc., Ashland, OR, USA).
3
Confocal Imaging of Fibrin Clot Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For laser scanning confocal microscopy, plasma was diluted 1/6 in tris-buffered saline, mixed with 50 µg/ml AlexaFluor488-labelled human fibrinogen, 0.1 U/ml human alpha thrombin (Merck, Millipore, Watford, UK) and 10 mM CaCl2 (final concentrations), and loaded into the channel of an uncoated µ-slide (Ibidi GmbH, Gräfelfing, Germany). Plasma clots were allowed to form for 2 h in dark humidity chambers followed by imaging using a Zeiss LSM880 with 40× and 63× oil immersion objectives (Carl Zeiss, Welwyn Carden City, UK). To determine if the aggregates contained cell-membrane or other hydrophobic structures, DiOC6(3) (3,3′-Dihexyloxacarbocyanine Iodide, Thermo Fisher, 1.2 µg/ml) was added prior to the addition of thrombin. To determine if aggregates were present prior to clotting, the same protocol was followed without adding thrombin. We collected z-stacks (20 µm, 30 slices) and images were projected to a single plane followed by analysis using ImageJ (Wayne Rasband, NIH).
4
Apoptosis Pathway Evaluation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AITC, 1,2-bis(2-aminophenoxy)ethane-N,N,N,N′-tetraacetic acid tetrakis(acetoxymethyl ester) (BAPTA/AM), calpeptin, 4′,6-diamidino-2-phenylindole (DAPI), N-acetylcysteine (NAC), propidium iodide (PI), and trypan blue were obtained from Sigma-Aldrich (Merck KGaA). 2′,7′-Dichlorodihydrofluorescein diacetate (H2DCFDA), 3,3′-dihexyloxacarbocyanine iodide [DiOC6(3 (link))], fetal bovine serum (FBS), 4-(6-acetoxymethoxy-2,7-dichloro-3-oxo-9-xanthenyl)-4′-methyl-2,2′(ethylenedioxy)dianiline-N, N,N,N′-tetraacetic acid tetrakis(acetoxymethyl) ester (fluo-3/AM), L-glutamine, Roswell Park Memorial Institute (RPMI)-1640 medium, penicillin, streptomycin, and trypsin-EDTA were obtained from Thermo Fisher Scientific, Inc. Caspase-3 and Caspase-9 Colorimetric Assay Kits were purchased from R&D Systems. Caspase-3 inhibitor Z-DEVD-FMK and caspase-9 inhibitor Z-LEHD-FMK were purchased from Merck Millipore. All primary antibodies were purchased from Cell Signaling Technology, Inc. or Santa Cruz Biotechnology, Inc. The goat anti-rabbit or anti-mouse immunoglobulin G (IgG) secondary antibodies conjugated with horseradish peroxidase (HRP) were also obtained from Cell Signaling Technology, Inc. Mitochondria/Cytosol Fractionation Kit was purchased from BioVision, Inc. The goat anti-mouse IgG-phycoerythrin (PE) secondary antibody was also obtained from Santa Cruz Biotechnology, Inc.
5
Simultaneous Mitochondrial and Membrane Labeling
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!