Fluorescein isothiocyanate (fitc)

Manufactured by Yeasen

Sourced in China

Fluorescein isothiocyanate (FITC) is a fluorescent dye commonly used in various laboratory techniques. It is a derivative of fluorescein and is known for its ability to emit green fluorescence when exposed to blue or ultraviolet light. FITC is commonly used for labeling and detecting proteins, cells, and other biomolecules in applications such as flow cytometry, immunohistochemistry, and fluorescence microscopy.

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7 protocols using fluorescein isothiocyanate (fitc)

Fluorescent FYGL Labeling and Cellular Uptake

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To detect the absorption of FYGL in cells, FITC-labeled FYGL was prepared. FITC
(Yeasen Biotechnology) was dissolved in DMSO, and water-soluble FYGL was dissolved in Na₂CO₃/NaHCO₃ buffer. FITC (1 mg/mL) was added into the FYGL solution (1 mg/mL) to form FITC-FYGL, and the mixture
was incubated in the dark at 4 °C for 12 h. Extensive dialysis
of the mixture was performed using 1 L of PBS in the dark until the
unreacted FITC was removed. Finally, FITC-FYGL was
obtained. The ultraviolet (UV) spectra of FITC, FYGL, and FITC-FYGL are shown in Figure S9. FITC has a maximum UV absorption at 495 nm, while FYGL is a proteoglycan and has a maximum UV absorption at
280 nm. It can be estimated that the ratio of FYGL and FITC in FITC-FYGL is 100:117 according to the
following equation:^{62 (link)} where F and P represent FITC
and protein, and A denotes the absorbance. C2C12 myoblasts were seeded
at a density of 2 × 10⁵ cells/well in six-well plates,
and fully differentiated myotubes were incubated with 100 μg/mL
FITC-FYGL for 4 h. DAPI (Beyotime, Shanghai, China)
and rhodamine phalloidin were used for nuclear and cytoskeleton staining,
respectively. The absorption of FITC-FYGL in C2C12
myotubes was observed under a laser confocal microscope (C2+; Nikon,
Tokyo, Japan).

Li J., Zhang Y., Yu F., Pan Y., Zhang Z., He Y., Yang H, & Zhou P. (2023). Proteoglycan Extracted from Ganoderma lucidum Ameliorated Diabetes-Induced Muscle Atrophy via the AMPK/SIRT1 Pathway In Vivo and In Vitro. ACS Omega, 8(33), 30359-30373.

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Doxorubicin-Loaded Mesoporous Silica Particles

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DOX was purchased from Melone Pharmaceutical Co., Ltd, Shanghai, China. Mesoporous silica particles (MSPs) with a diameter of 1 μm were purchased from Huake Microtechnology Co., Ltd, Wuhan, China. Lysine was purchased from Aladdin Biochemical Technology Co., Ltd, Shanghai, China. FITC was purchased from Yeasen Biotechnology Co., Ltd, Shanghai, China. To load DOX, 50 mg of DOX was dissolved in 5 mL deionized water (DI) at a mass concentration of 10 mg/mL. Then, 100 mg MSP was added to 5 mL DOX solution. It was stirred for 24 hours at 25 °C in a shaker at 180 rpm. Finally, it was centrifuged at 2000 rpm for 10 min, and the supernatant was removed to obtain a DLSP solution. Lysine was diluted tenfold, and 5 mL diluted lysine was added to the DLSP sample. The sample was stirred for 6 h on a magnetic stirrer. Positively charged 1-μm mesoporous silica particles loaded with DOX were obtained. The DLSP pellets were placed in a 1 mL centrifuge tube. At the same time, FITC with a mass concentration of 1 mg/mL was added to the centrifuge tube. The mixture was incubated at room temperature for 4 h in the dark. Finally, a green fluorescent DLSP was obtained.

Xiong J., Li X., He Z., Shi Y., Pan T., Zhu G., Lu D, & Xin H. (2024). Light-controlled soft bio-microrobot. Light, Science & Applications, 13, 55.

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Doxorubicin Nanoparticle Cytotoxicity Assay

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Doxorubicin hydrochloride (DOX) was purchased from Sangon (Shanghai, China). Calcium chloride (CaCl 2 , 96%), disodium hydrogen phosphate (Na 2 HPO 4 , 99%) were obtained from Titan (Shanghai, China). Dopamine hydrochloride and 2-Amino-2-(hydroxymethyl)-1,3-propanediol (Tris) were obtained from Biofroxx (Guangzhou, China). FITC was purchased from Yeasen (Shanghai, China). Bovine serum albumin (BSA), 4′,6-diamidino-2-phenylindole (DAPI), 70 µm cell screen mesh, Live/Dead Viability/Cytotoxicity Kit (Promega), Advanced DMEM/F12 (Gibco), B-27 Supplement(50X), N2-supplement (100X), N-acetyl-L-cysteine, Recombinant Mouse EGF were purchased from Gibco (USA). Puromycin, Zeocin, Plasmocin, trypsin-ethylenediaminetetraacetic acid (trypsin-EDTA) were gained from Invitrogen (USA). Matrigel Matrix was obtained from Corning (USA). CS10 was obtained from Stem Cell (Canada). CellTiter-Glo 3D Cell Viability Assay was purchased from Promega (USA). Rhodamine-phalloidin was purchased from Thermo Fisher (USA).

Deng T., Luo D., Zhang R., Zhao R., Hu Y., Zhao Q., Wang S., Iqbal M.Z, & Kong X. (2023). DOX-loaded hydroxyapatite nanoclusters for colorectal cancer (CRC) chemotherapy: Evaluation based on the cancer cells and organoids. SLAS technology, 28(1).

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Hydrothermal Synthesis of Functionalized HNTs

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HNTs were provided by Guangzhou Runwo Materials Technology Co., Ltd., China and used without further purification. The elemental composition of used HNTs by X-ray fluorescence (XRF) was determined as follows (wt%): SiO₂, 58.91; Al₂O₃, 40.41; Fe₂O₃, 0.275; P₂O₅, 0.138; TiO₂, 0.071. The weight loss of HNTs was 19.20%, which was mainly occurred in the temperature range of 400–500 °C. Fluorescein isothiocyanate (FITC) (analytical grade) was purchased from Shanghai Yeasen Biotechnology Co., Ltd., Shanghai, China. Indocyanine green (ICG) and streptavidin (SA) were purchased from Sigma-Aldrich (St. Louis, MO, US). Anti-EpCAM aptamer was purchased from Sangon Biotech, Shanghai. Acridine orange (AO) and ethidium bromide (EB) were purchased from Solarbio Science and Technology Co., Ltd., Beijing, China. Ultrapure water was obtained from Milli-Q water system. All the other chemicals were analytically graded and used directly without further purification.

Tan C., Zheng J., Feng Y, & Liu M. (2021). Cell Membrane-Coated Halloysite Nanotubes for Target-Specific Nanocarrier for Cancer Phototherapy. Molecules, 26(15), 4483.

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Synthesis of Fluorescent Silica Nanoparticles

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Tetraethyl orthosilicate (TEOS), ammonium hydroxide (NH₄OH, 25-28%), ethanol (> 99%), cetyltrimethylammonium bromide (CTAB), sodium carbonate (Na₂CO₃), sodium hydroxide (NaOH), hydrochloric acid (HCl, 36-38%), 3-aminopropyl triethoxysilane (APTES), toluene (> 99%), methanol (> 99%), calcium peroxide (CaO₂) and polyacrylic acid (PAA, M.W. ~3000) were purchased from Sinopharm Chemical Reagent Co., Ltd. (China). Fluorescein isothiocyanate (FITC) was obtained from Yeasen Biotech Co., Ltd. (China). Phosphate-buffered saline (PBS, without calcium and magnesium) was acquired from Corning Incorporated (USA). All reagents were used as received without further purification. The deionized water used in all experiments was produced by a Milli-Q water purification system with a specific resistance greater than 18 MΩ.cm at 25 °C.

Wu D., Zhu Z.Q., Tang H.X., Shi Z.E., Kang J., Liu Q, & Qi J. (2020). Efficacy-shaping nanomedicine by loading Calcium Peroxide into Tumor Microenvironment-responsive Nanoparticles for the Antitumor Therapy of Prostate Cancer. Theranostics, 10(21), 9808-9829.

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Multifunctional Nanoparticle Delivery System

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Polyethylene glycol (PEG, MW = 4.0 kDa) and GSH were purchased from Sigma–Aldrich. Succinimide ester-polyethylene glycol₅₀₀₀-cyclo (Asp-d-Phe-Lys-Arg-Gly) (NHS-PEG₅₀₀₀-cRGD) was purchased from Beijing Jenkem Technology Co., Ltd. CPT was purchased from Dalian Meilun Biology Technology Co., Ltd. Citric acid monohydrate, triphosgene, K₄[Fe(CN)₆]·3H₂O, FeCl₃·6H₂O, polyallylamine hydrochloride (PAH, MW = 15.0 kDa), and 2,2′-dithiodiethanol were purchased from Shanghai Aladdin Chemistry Co., Ltd. HA (MW = 32.0 kDa) was purchased from Bloomage Biology Technology Co., Ltd. HSP70 antibody was purchased from Abcam. Lyso-Tracker red and 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) were purchased from Beyotime Biotechnology Co., Ltd. Calcein-AM/propidium iodide (Calcein-AM/PI) Double Stain Kit, fluorescein isothiocyanate (FITC), and Annexin V-FITC/PI Apoptosis Detection Kit were purchased from Shanghai Yeasen Biotechnology Co., Ltd. Murine 4T1 breast cancer cells were obtained from Chinese Academy of Sciences Cells Bank (Shanghai, China). All the materials used in this study were analytic grade and used without further purification.

Yang Y., Hu D., Lu Y., Chu B., He X., Chen Y., Xiao Y., Yang C., Zhou K., Yuan L, & Qian Z. (2021). Tumor-targeted/reduction-triggered composite multifunctional nanoparticles for breast cancer chemo-photothermal combinational therapy. Acta Pharmaceutica Sinica. B, 12(6), 2710-2730.

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Optimized Core-Shell Nanoparticle Formulation

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The PFTBA core solution preparation was carried out in accordance with previous investigations [33 (link)]. Briefly, 190 mg of egg yolk lecithin (Sigma, USA) was dissolved in 1 mL of Tyrode's solution (Sigma, USA), and mixed via ultrasonic shaker twice at 4 °C for 15 s. Subsequently, 1 mL of PFTBA stock solution was introduced, followed by sonic concussion 10 times under the same parameters. In control groups (without PFTBA), PBS (Gibco, USA) was introduced instead. Finally, carboxyl-chitosan (Sigma, USA) was introduced to the above emulsion to form a gelatinized core solution. Varying concentrations of carboxyl chitosan (5% w/v, 7.5% w/v, 10% w/v, 12.5% w/v, 15% w/v; 20% w/v, Sigma, USA) were used to form the optimal structure. In addition, to prepare the shell solution, 20% w/v polycaprolactone (PCL, Sigma, USA) particles were dissolved in methanol-chloroform solution, in a 1:4 v/v ratio. To observe the electrospinning process, Rhodamine B (1 mg/mL, Sigma, USA) was mixed with the shell material, fluorescein isothiocyanate (FITC; 2 mg/mL, YEASEN, China) with the core material, and the core-shell structure was fluorescently labeled during the electrospinning process.

Zheng Y., Xue B., Wei B., Xia B., Li S., Gao X., Hao Y., Wei Y., Guo L., Wu H., Yang Y., Gao X., Yu B., Zhang Y., Yang S., Luo Z., Ma T, & Huang J. (2023). Core-shell oxygen-releasing fibers for annulus fibrosus repair in the intervertebral disc of rats. Materials Today Bio, 18, 100535.

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