Cy3 azide

Manufactured by Lumiprobe

Sourced in Uruguay

Cy3-Azide is a fluorescent dye molecule that can be used for labeling biomolecules such as proteins, nucleic acids, or small molecules. It contains a Cy3 fluorescent group and an azide functional group, which allows for the dye to be conjugated to target molecules through click chemistry reactions.

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Lab products found in correlation

Cy5.5 nhs dye, by Lumiprobe (1 mentions) L ascorbic acid, by Merck Group (1 mentions) Paclitaxel ptx, by LC Laboratories (1 mentions) Stannous octoate, by MP Biomedicals (1 mentions) Alexa fluor 647 azide, by Vector Laboratories (1 mentions) Alexafluor488 azide, by Vector Laboratories (1 mentions) 5 ethynyl 2 deoxyuridine (edu), by Lumiprobe (1 mentions)

4 protocols using cy3 azide

Synthesis of Paclitaxel-Loaded Polymeric Micelles

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Methoxy poly(ethylene oxide) (PEO) (M_wt 5000 Da), sodium dodecyl sulfate, and L-Ascorbic
acid (99%) were purchased from Sigma (St Louis, MO). ε-Caprolactone
was purchased from Lancaster synthesis (U.K.). α-Benzylcarboxylate-ε-caprolactone
(BCL) and α-propargyl-carboxylate-ε-caprolactone were
synthesized by Alberta Research Chemicals Inc. (ARCI, Edmonton, AB,
Canada) based on methods published previously by our group.^{39 (link)} 1, 2-Distearoyl-sn-glycero-3-phosphoethanolamine-N-[succinimidyl (polyethylene glycol)] (DSPE-PEG3400-NHS)
and methoxy poly(ethylene oxide) (M_wt 2000
Da), covalently linked to distearoyphosphadidyl ethanolamine (mPEO2000-DSPE),
were purchased from Nanocs Inc. Paclitaxel (PTX) (purity> 99.5)
was
purchased from LC Laboratories (Woburn, MA). Rituximab (anti CD20
antibody) and Taxol were provided by Cross Cancer Institute. Stannous
octoate was purchased from MP Biomedicals Inc. (Germany). Spectra/por
dialysis tubing (MWCO, 3.5kDa) was purchased from Spectrum Laboratories
(Rancho Dominguez, CA). Cy3 azide, Cy5.5 NHS dye, and Cu(II) (Tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine) (TBTA) complex were purchased
from Lumiprobe (Hallandale Beach, Florida). All other chemicals and
reagents used were of analytical grade.

Saqr A., Vakili M.R., Huang Y.H., Lai R, & Lavasanifar A. (2019). Development of Traceable Rituximab-Modified PEO-Polyester Micelles by Postinsertion of PEG-phospholipids for Targeting of B-cell Lymphoma. ACS Omega, 4(20), 18867-18879.

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EdU Labeling and Immunostaining Protocol

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EdU was given by intraperitoneal injection at a concentration of 5 µg g⁻¹. For EdU staining, sections were permeabilized by incubation for 30 min in 0.5% Triton X-100 in PBS followed by 3 × 5-min washes in PBS. EdU staining solution was made fresh each time, containing 100 mM Tris, 4 mM CuSO₄, and 100 mM ascorbic acid diluted in 1× PBS. The fluorescently labeled azide molecule was added last, and the staining cocktail was immediately added to sections for a 30-min incubation at room temperature (final concentrations of each fluorescent molecule: 4 mM AlexaFluor488-Azide [Click Chemistry Tools, 1275-1], 10 mM Cy3-Azide [Lumiprobe, A1330], 16 mM AlexaFluor647-Azide [Click Chemistry Tools, 1299-1]). After incubation, sections were washed 3 × 5 min in PBS. After EdU labeling, standard protocols were followed for immunostaining.

Keil J.M., Doyle D.Z., Qalieh A., Lam M.M., Funk O.H., Qalieh Y., Shi L., Mohan N., Sorel A, & Kwan K.Y. (2020). Symmetric neural progenitor divisions require chromatin-mediated homologous recombination DNA repair by Ino80. Nature Communications, 11, 3839.

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Visualizing DNA Synthesis in HeLa Cells

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HeLa cells are cultured according to standard procedures and seeded 1 or 2 days before the treatment on 12 mm diameter #1 glass coverslips. HeLa cells are incubated with 10 μM EdU (5-Ethynyl-2′-deoxyuridine, Lumiprobe) for 30 minutes at 37°C. The cells are fixed with 4% formaldehyde in PBS and permeabilised with 0.1% Triton X-100 in PBS. Click chemistry is performed with 9 μM Cy3-azide (Lumiprobe) and 2 mM CuSO₄. To start the reaction, 20 mg/ml ascorbate (final concentration) is added, and the solution is used immediately to stain the cells. After 30 minutes, the cells are washed 3× with PBS, and the sample is incubated with 0.1 μg/ml DAPI for 5 minutes. Samples are mounted in Mowiol (24% glycerol (w/v), 0.1 g mL–1 polyvinylalchol 4.88, 0.1 M Tris-HCl (pH 8.5) in H2O) and used for observation with fluorescence microscopy.

, & Goedhart J. (2024). Studentsourcing—Aggregating and reusing data from a practical cell biology course. PLOS Computational Biology, 20(2), e1011836.

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Synthesis of Cy3-Labeled Macromonomer

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Example 11

Cy3-MM was prepared in an analogous fashion to that reported for Cy5.5-MM. Briefly, 13.4 mg of Cy3-Azide (Lumiprobe) was added to a 2-dram vial. Next, 60 mg of an alkyne-containing macromonomer¹¹were added. The combined solids were transferred into a nitrogen-filled glovebox and dissolved in 1 mL of DCM. Next, a spatula tip of copper (I) acetate was added and the solution stirred for 1 h, after which time LC-MS analysis indicated full conversion of Cy3-Azide. The mixture was then concentrated and purified by preparative HPLC using an acetonitrile-0.1% AcOH in water gradient. The desired fractions were partially concentrated by rotary evaporation until ˜25% of the residual solvent remained, then 250 mL of DCM were added and the entire mixture was dried with sodium sulfate. The desired product was collected by removal of the sodium sulfate and concentration to yield Cy3-MM as a magenta solid. Calculated mass (M)⁺: 3698.84 found 3696.08 (see FIGS. 63A-63B).

US10988491B2. Degradable polymers of a cyclic silyl ether and uses thereof (2021-04-27). Massachusetts Institute of Technology [US]. Inventors: Jeremiah A. Johnson [US], Peyton Shieh [US], Wenxu Zhang [US].

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