Fluorescent dyes

Manufactured by Merck Group

Sourced in Germany

Fluorescent dyes are a class of chemical compounds that emit light when exposed to specific wavelengths of light. They are widely used in various scientific applications, including cell biology, molecular biology, and analytical chemistry, to label and visualize specific biological structures or molecules.

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Fluorescent dyes (1,1′-dioctadecyl-3,3,3′,3′ tetramethylindocarbocyanine perchlorate (DiI), N-Hydroxysuccinimide (NHS), Lysotracker DCFH-DA fluorescent dyes PKH26 red fluorescent dye JC‐1 cationic fluorescent dye kit 7-Azido-4-Methylcoumarin (AzMC) fluorescent dye PKH26 Red Fluorescent cell dye Cy3 fluorescent dye SYBR-Green Fluorescent dye Nile red fluorescent dye DCFDA fluorescent dye

3 protocols using fluorescent dyes

Mass Spectrometry and HPLC Analysis

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Reagents and solvents were purchased from Sigma Aldrich (Taufkirchen, Germany), Glen Research (Sterling, VA, USA), or Alfa Aesar (Thermo Fisher Scientific, Waltham, MA, USA) and were used without further purification. Phospholipids were purchased from Bachem AG (Bubendorf, Switzerland) or Avanti Polar Lipids (Alabaster, AL, USA). Fluorescent dyes were purchased from Sigma Aldrich (Germany). Cell culture media and additives were from Gibco^® (Thermo Fisher Scientific, Waltham, MA, USA).
For the mass spectrometry analysis, an Ultraflex III MALDI-TOF-TOF instrument (Bruker Daltonics GmbH, Bremen, Germany) equipped with a Smartbeam 2 laser with a repetition rate up to 200 Hz (500 laser shots) was used. The obtained mass spectrum was analyzed with Flex-analysis version 4.0 software (Bruker Daltonics GmbH).
The HPLC separation and analysis were performed with Waters^® Alliance 2695 (Waters^® Corporation, Milford, MA, USA) apparatus equipped with the ultraviolet (UV) detector Waters^® 486 (Waters Corporation) and analyzed with Millennium 32 software (Waters Corporation). The reverse phase analytical C8 column (Macherey-Nagel, GmbH & Co, KG, Hoerdt, France) was used for all separations.

Beztsinna N., Tsvetkova Y., Jose J., Rhourri-Frih B., Al Rawashdeh W., Lammers T., Kiessling F, & Bestel I. (2017). Photoacoustic imaging of tumor targeting with riboflavin-functionalized theranostic nanocarriers. International Journal of Nanomedicine, 12, 3813-3825.

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Synthesis of Fluorescent Lipid Nanoparticles

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CIP, potassium hydroxide (KOH), acetic anhydride, DMSO, DCM, sodium cyanoborohydride (NaBH₄), oils, surfactants, co-surfactants, pluronic 127, culture media, fluorescent dyes, and Schiff reagent were obtained from Sigma-Aldrich, Germany. All solvents used were of analytical grade.

Arshad R., Arshad M.S., Tabish T.A., Shah S.N., Afzal S, & Shahnaz G. (2022). Amidated Pluronic Decorated Muco-Penetrating Self-Nano Emulsifying Drug Delivery System (SNEDDS) for Improved Anti-Salmonella typhi Potential. Pharmaceutics, 14(11), 2433.

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Embryonic Development Morphology and Permeability

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The morphology of embryos was examined by using embryos dissected from day 1 adults followed by mounting on a 2% agarose pad on a glass slide. DIC and fluorescent images were taken by using Leica DM 2500 (Leica, Ernst-Leitz-Straße, Wetzlar, Germany). For the osmotic sensitivity assay, embryos were placed in water, egg buffer or 1 M potassium chloride for 1 h at room temperature. For the dye permeability assay, embryos were stained with 0.4% trypan blue or 10 g/ml fluorescent dyes (Sigma-Aldrich) in PBS in the dark for 30 min. For the permeability barrier assay, embryos in the genetic background of OD344 (mCherry::CPG-2; GFP::PH) were used. The integrity of the permeability barrier was determined by the localization of mCherry::CPG-2. To quantify polarity, the area of two daughter cells was analyzed by Image J (NIH, Bethesda, MD, USA). The polarity defect was defined as the area of the anterior cell less than 53% of total embryonic cells. For brood size measurement, the total number of viable F1 worms derived from RNAi-treated parents was determined. For the embryonic development determination, time-lapsed images of embryos were taken at room temperature by using Leica DM 2500 (Leica).

Chen T.L., Yang H.C., Hung C.Y., Ou M.H., Pan Y.Y., Cheng M.L., Stern A., Lo S.J, & Chiu D.T. (2017). Impaired embryonic development in glucose-6-phosphate dehydrogenase-deficient Caenorhabditis elegans due to abnormal redox homeostasis induced activation of calcium-independent phospholipase and alteration of glycerophospholipid metabolism. Cell Death & Disease, 8(1), e2545-.

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