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Diic16

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DiIC16 is a fluorescent dye used for labeling and tracking cell membranes in biological samples. It is a long-chain dialkylcarbocyanine dye that inserts into the lipid bilayer of cell membranes, providing a stable and specific staining of the cell surface.

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Lab products found in correlation

Goat antihuman, by LI COR (2 mentions) Goat anti mouse igg, by LI COR (2 mentions) Mtz nhs, by Vector Laboratories (2 mentions) Dii c18 2, by Thermo Fisher Scientific (2 mentions) Dii c18, by Biotium (2 mentions) Goat igg fraction to mouse c3 and human c3 catalog no 55 463, by MP Biomedicals (2 mentions) Chrompure human and mouse igg, by Jackson ImmunoResearch (2 mentions) Irdye 80 cw labeled donkey antigoat, by LI COR (2 mentions) Dii c12, by Thermo Fisher Scientific (2 mentions) Fatty acids, by Tokyo Chemical Industry (2 mentions) Diic16, by Agilent Technologies (1 mentions) Histodenz, by Merck Group (1 mentions) Synergy 4, by Agilent Technologies (1 mentions) Bx50 epifluorescence microscope, by Olympus (1 mentions) Dharmafect 1, by Thermo Fisher Scientific (1 mentions) Polarstar omega plate reader, by BMG Labtech (1 mentions)

Close spelling variants of this product

1,1’-dihexadecyl-3,3,3’,3’-tetramethylindocarbocyanine perchlorate (DiIC16(3);

6 protocols using diic16

1

Exosomes Labeling and Uptake

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To prepare DiIC16-labeled exosomes, QGY-7703 cells were washed twice with 1× PBS 24 hr after seeding, followed by incubation for 48 hr in the serum-free DMEM containing DiIC16 (cat. D384; Invitrogen, Carlsbad, CA, USA) at a final concentration of 1 μg/mL. The DiIC16-containing medium that was incubated without QGY-7703 cells was used as a control for detecting any carryover of free DiIC16. The medium was then applied to sequential ultracentrifugation to collect exosomes as described above.
To prepare exosomes from Cy3-labeled miR-210 transfectants, QGY-7703 cells transfected with 50 nM Cy3-labeled miR-210 (Cy3-miR-210) were washed as above, followed by replacement with 10 mL 10% FBS-containing DMEM for 48 hr. The exosomes were collected by sequential ultracentrifugation as described above.
HUVEC cells were incubated with exosomes in 1% FBS-containing SFM for 24 hr and then washed with 1× PBS thrice before imaging under a fluorescence microscope.
Lin X.J., Fang J.H., Yang X.J., Zhang C., Yuan Y., Zheng L, & Zhuang S.M. (2018). Hepatocellular Carcinoma Cell-Secreted Exosomal MicroRNA-210 Promotes Angiogenesis In Vitro and In Vivo. Molecular Therapy. Nucleic Acids, 11, 243-252.
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2

Caveolin Protein Density Profiling

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GFP-tagged caveolin WT and truncation mutants were expressed (75 µl each) in the presence of a lipid dye, DiIC16 (D384; Invitrogen; at 1:1,000 dilution), and mixed with an equal volume of 80% (wt/vol) Histodenz (Sigma-Aldrich) solution in PBS. The mixed samples were loaded at the bottom of 11 × 22 mm Beckman centrifuge tubes (Beckman Instrument). The tube was sequentially overlaid with a linear Histodenz gradient (7.5%–37.5% wt/vol) in PBS. Gradients were spun at 150,000 g for 16 h in a TLS-55 rotor (Beckman Instrument) at 4°C. After that, samples were collected from the top in 50-µl increments and loaded into 384-well white plates. Fluorescence levels of GFP and DiIC16 for each fraction were read on a plate reader (Synergy 4; BioTek; excitation 485 nm and emission 520 nm for GFP and 549 nm/575 nm for DiIC16, respectively).
Jung W., Sierecki E., Bastiani M., O’Carroll A., Alexandrov K., Rae J., Johnston W., Hunter D.J., Ferguson C., Gambin Y., Ariotti N, & Parton R.G. (2018). Cell-free formation and interactome analysis of caveolae. The Journal of Cell Biology, 217(6), 2141-2165.
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3

EGCG and Luteolin Modulate Fibroblast Morphology

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WPMY-1 fibroblasts were plated at 50% confluency. The following day, the cells were treated with 5–40 µM EGCG or luteolin +/−5 ng/ml TGF-β for 24 hours. DiIC16 (Molecular Probes, Invitrogen) was added directly to the media at a final concentration of 1 µM and incubated at room temperature for 2 minutes. Cells were fixed with 4% paraformaldehyde, washed and nuclei stained with DAPI. Images were captured on an Olympus BX-50 epifluorescence microscope using MetaMorph software.
Gray A.L., Stephens C.A., Bigelow R.L., Coleman D.T, & Cardelli J.A. (2014). The Polyphenols (−)-Epigallocatechin-3-Gallate and Luteolin Synergistically Inhibit TGF-β-Induced Myofibroblast Phenotypes through RhoA and ERK Inhibition. PLoS ONE, 9(10), e109208.
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4

MYO1C Depletion Impacts Cell Migration

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MCF10A cells were seeded in 6-well plates and transfected after 24 hours with either scrambled siRNA, serving as a control, or Dharmacon siRNA for MYO1C (M-015121-005, Thermo Scientific) to a final concentration of 20 μM (as recommended by the manufacturer) using DharmaFECT1 (Thermo Scientific). After incubation for 24 hours, cells were trypsinized, counted, diluted to 2x105 cells per ml and incubated with 2.5 μM DiI-C16 (Molecular Probes, Invitrogen) for 30 min at 37°C, washed and 4x104 cells per well were plated in the OrisTM Cell Migration Assay plates (Platypus Technologies) containing cell seeding stopper. Cells were allowed to attach and spread overnight at 37°C before the stoppers were removed and medium was refreshed. Migration was measured in real time using the POLARstar Omega plate reader (BMG LABTECH, Germany) with temperature at 37°C and 5% CO2 collecting data points every 20 min for up to 24 hours. Migration speed was measured by calculating the area under the curve for each well. Pictures were taken before and after the migration assays. Each treatment was done in triplicate.
Visuttijai K., Pettersson J., Mehrbani Azar Y., van den Bout I., Örndal C., Marcickiewicz J., Nilsson S., Hörnquist M., Olsson B., Ejeskär K, & Behboudi A. (2016). Lowered Expression of Tumor Suppressor Candidate MYO1C Stimulates Cell Proliferation, Suppresses Cell Adhesion and Activates AKT. PLoS ONE, 11(10), e0164063.
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5

Lipid Labeling for Cellular Imaging

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Fatty acids were from TCI America (Portland, OR, USA), PEG was from Laysan Bio (Arab, AL, USA), and MTz-NHS was from Click Chemistry Tools. DiI-C18:2, and DiI-C16, DiI-C12 were from ThermoFisher (Waltham, MA, USA). DiI-C18 (1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindotricarbocyanine Perchlorate) and DiR (1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindotricarbocyanine Iodide) were from Biotium (Hayward, CA, USA). DiI-NH2 was synthesized by a previously reported method.[10 (link)] All the lipids were stored as 1 mM stocks in ethanol. Anticoagulant Citrate Dextrose (ACD) buffer was obtained from the Children’s Hospital Colorado Blood Donation Center and kept sterile before use. Goat IgG fraction to mouse C3 and human C3 (Catalog No. 55 463 and 55 033, respectively) was from MP Biomedicals (Solon, OH, USA). ChromPure human and mouse IgG was from Jackson Immuno Research (West Grove, PA, USA). Secondary antibodies IRDye 80°CW labeled donkey antigoat, goat antihuman, and goat antimouse IgG were from LI-COR Biosciences (Lincoln, NE, USA).
Gaikwad H., Wang G., Li Y., Bourne D, & Simberg D. (2022). Surface Modification of Erythrocytes with Lipid Anchors: Structure–Activity Relationship for Optimal Membrane Incorporation, in vivo Retention, and Immunocompatibility. Advanced nanobiomed research, 2(8), 2200037.
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6

Lipid Labeling for Cellular Imaging

Cited 1 time
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Fatty acids were from TCI America (Portland, OR, USA), PEG was from Laysan Bio (Arab, AL, USA), and MTz-NHS was from Click Chemistry Tools. DiI-C18:2, and DiI-C16, DiI-C12 were from ThermoFisher (Waltham, MA, USA). DiI-C18 (1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindotricarbocyanine Perchlorate) and DiR (1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindotricarbocyanine Iodide) were from Biotium (Hayward, CA, USA). DiI-NH2 was synthesized by a previously reported method.[10 (link)] All the lipids were stored as 1 mM stocks in ethanol. Anticoagulant Citrate Dextrose (ACD) buffer was obtained from the Children’s Hospital Colorado Blood Donation Center and kept sterile before use. Goat IgG fraction to mouse C3 and human C3 (Catalog No. 55 463 and 55 033, respectively) was from MP Biomedicals (Solon, OH, USA). ChromPure human and mouse IgG was from Jackson Immuno Research (West Grove, PA, USA). Secondary antibodies IRDye 80°CW labeled donkey antigoat, goat antihuman, and goat antimouse IgG were from LI-COR Biosciences (Lincoln, NE, USA).
Gaikwad H., Wang G., Li Y., Bourne D, & Simberg D. (2022). Surface Modification of Erythrocytes with Lipid Anchors: Structure–Activity Relationship for Optimal Membrane Incorporation, in vivo Retention, and Immunocompatibility. Advanced nanobiomed research, 2(8), 2200037.
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