Fluorescein isothiocyanate dextran fitc dextran

Manufactured by Merck Group

Sourced in United States, Germany

Fluorescein isothiocyanate-dextran (FITC-dextran) is a fluorescent labeling agent used in various research applications. It consists of the fluorescent compound fluorescein isothiocyanate (FITC) covalently attached to dextran, a branched polysaccharide. FITC-dextran can be used to visualize and track the movement of molecules, cells, or other biological entities in experimental settings.

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Lab products found in correlation

Chloroform, by Merck Group (2 mentions) Inverted microscope, by Zeiss (2 mentions) P stat3, by Cell Signaling Technology (1 mentions) Human il 4, by Thermo Fisher Scientific (1 mentions) Human il 13, by Thermo Fisher Scientific (1 mentions) Anti cd163, by Santa Cruz Biotechnology (1 mentions) Anti cd206, by Santa Cruz Biotechnology (1 mentions) Anti β2 ar, by Santa Cruz Biotechnology (1 mentions) Anti pka, by Santa Cruz Biotechnology (1 mentions) Anti creb antibody, by Abcam (1 mentions) Alexa fluor 488 donkey anti mouse antibody, by Thermo Fisher Scientific (1 mentions) Alexa fluor 594 goat anti rabbit antibody, by Thermo Fisher Scientific (1 mentions) Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions) Phenylmethylsulfonyl fluoride pmsf, by Beyotime (1 mentions) Fitc dextran, by Thermo Fisher Scientific (1 mentions) Transwell clear insert, by Corning (1 mentions) Polycaprolactone pcl, by Merck Group (1 mentions) Polylactic acid pla, by Merck Group (1 mentions) Dimethyl itaconate, by Merck Group (1 mentions) Barnstead nanopure water purification system, by Thermo Fisher Scientific (1 mentions)

27 protocols using fluorescein isothiocyanate dextran fitc dextran

Macrophage Polarization Pathway Analysis

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Phorbol-12-myristate-13-acetate (PMA) was bought from Sigma Aldrich (St. Louis, Missouri, USA), human IL-4 and human IL-13 were purchased from Peprotech (200-04 and 200-13, Rocky Hill, USA).The anti-CD163, anti-CD206, anti-β2-AR and anti-PKA antibodies were purchased from Santa Cruz Biotechnology(sc-33559, sc-58986, sc-271322 and sc-98951, Ca, USA). Phenylmethylsulfonyl fluoride (PMSF) was bought from beyotime biotechnology (Shanghai, China). Primary antibodies against STAT3, pSTAT3, pCREB were purchased from Cell Signaling Technology (4904s, 9131s and 9198s, Danvers, MA, USA). Anti-CREB antibody was purchased from Abcam (ab32515, Cambridge, MA, USA). AlexaFluor488 donkey anti-mouse antibody and AlexaFluor594 goat anti-rabbit antibody were purchased from ThermoFisher Scientific (A11037, A21202, Waltham, MA, USA). Fluorescein isothiocyanate dextran (FITC-dextran) was from Sigma Aldrich (FD40s, St. Louis, Missouri, USA). AlexFluor 647 was from Fcmacs (FMS-Msaf64701, Nanjing, Jiangsu, China).

Wu J.J., Yang Y., Peng W.T., Sun J.C., Sun W.Y, & Wei W. (2019). G protein-coupled receptor kinase 2 regulating β2-adrenergic receptor signaling in M2-polarized macrophages contributes to hepatocellular carcinoma progression. OncoTargets and therapy, 12, 5499-5513.

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Transwell Epithelial Permeability Assay

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Primary AECs cells (n = 2 donors) were cultured on the apical side of Transwell® Clear inserts (Corning, VWR, Dublin, Ireland) until the formation of a confluent epithelial monolayer and treated with conditioned media (CM) of IPFFs (n = 3 patients) replete with, or depleted of, SPARC. After 6 days of CM treatment, 4 kDa dextran labeled with fluorescein isothiocyanate–dextran (FITC-dextran, 2 mg/ml) (Sigma-Aldrich, Dublin, Ireland) was added to the apical compartment of the tranwell culture and after 24 h the fluorescence signal from the FITC-dextran passed across the epithelial monolayer in the basal compartment was detected using a Fluoroskan plate reader Ascent FL (Thermofisher, Basingstoke, UK).

Conforti F., Ridley R., Brereton C., Alzetani A., Johnson B., Marshall B.G., Fletcher S.V., Ottensmeier C.H., Richeldi L., Skipp P., Wang Y., Jones M.G, & Davies D.E. (2020). Paracrine SPARC signaling dysregulates alveolar epithelial barrier integrity and function in lung fibrosis. Cell Death Discovery, 6, 54.

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Fluorescent Detection of Serotonin Release

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2,4-dinitrophenylated albumin from bovine serum (DNP-BSA) was purchased from Life Technologies (Carlsbad, CA). Monoclonal anti-Dinitrophenyl (anti-DNP) IgE, fluorescein isothiocyanate-dextran (FITC-dextran) and serotonin molecules were purchased from Sigma-Aldrich (St. Louis, MO). All solutions used in the experiments were prepared with home-made fresh extracellular buffer (135 mM NaCl, 5 mM KCl, 20 mM HEPES, 1.8 mM CaCl₂, 1 mM MgCl₂, 5.6 mM glucose, pH = 7.4). All reagents were analytical grade from Sigma-Aldrich, except those stated.

Zhang F., Guan Y., Yang Y., Hunt A., Wang S., Chen H.Y, & Tao N. (2019). Optical tracking of nanometer-scale cellular membrane deformation associated with single vesicle release. ACS sensors, 4(8), 2205-2212.

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Development of Therapeutic Tracheal Stent

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Poly (D,L-lactide-co-glycolide) (PLGA)—lactide:glycolide (50:50), Mw ∼30,000–60,000; polycaprolactone (PCL)—Mw ∼80,000; polylactic acid (PLA)—Mw ∼60,000; fluorescein isothiocyanate-dextran (FITC-dextran) Mw ∼5000, and chloroform were obtained from Sigma-Aldrich, Germany. Anti-inflammatory drug—methylprednisolone sodium succinate (Solu-Medrol) was obtained from Pfizer, Belgium. The Poly (dimethylsiloxane) (PDMS) kit (Sylgard 184) was purchased from Dow-Corning, Midland, USA. DexStent-TN (nitinol tracheal stents, d = 8 mm, l = 60 mm; and delivery systems, d = 10 Fr, l = 470 mm) were obtained from Dextronix, France.

Sindeeva O.A., Prikhozhdenko E.S., Schurov I., Sedykh N., Goriainov S., Karamyan A., Mordovina E.A., Inozemtseva O.A., Kudryavtseva V., Shchesnyak L.E., Abramovich R.A., Mikhajlov S, & Sukhorukov G.B. (2021). Patterned Drug-Eluting Coatings for Tracheal Stents Based on PLA, PLGA, and PCL for the Granulation Formation Reduction: In Vivo Studies. Pharmaceutics, 13(9), 1437.

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Synthesis and Characterization of Fluorescent Polymeric Nanoparticles

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1,4-dioxane, citric acid (CA), dimethyl itaconate (DMI), fluorescein isothiocyanate-dextran (FITC-dextran, MW: 2,000,000), polysucrose 70-fluorescein isothiocyanate conjugate (FITC-Ficoll 70), maleic anhydride (MA), 1,8-octanediol (OD), triethylene citrate (TEC), 4-methoxyphenol (MeHQ), Pluronic F127, stannous octanoate (SnOct), rhodamine B were all purchased from Sigma Aldrich (Oakville, ON). chloroform (used in post-synthesis cleaning) and methanol were purchased from BioShop Canada Inc. (Burlington, ON). Deionized water (DIW) was generated using a Thermo Scientific Barnstead NANOpure water purification system.

Liu C., Campbell S.B., Li J., Bannerman D., Pascual-Gil S., Kieda J., Wu Q., Herman P.R, & Radisic M. (2022). High throughput omnidirectional printing of tubular microstructures from elastomeric polymers. Advanced healthcare materials, 11(23), e2201346.

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Perfluorocarbon Emulsion Protocol

Cited 1 time

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1-oleoyl (C_18:1) LPA and fluorescein isothiocyanate–dextran (FITC-dextran; 4 kDa) were purchased from Sigma–Aldrich (Milwaukee, WI, USA). The alamar blue cell viability reagent was purchased from Life Technologies (Carlsbad, CA, USA). Bovine serum albumin (BSA) was purchased from New England Biolabs (Ipswich, MA, USA). Crystal violet (0.1% aqueous) was purchased from Ward’s Science (Rochester, NY, USA). Luria-Bertani (LB) agar powder was purchased from ThermoFisher Scientific (Waltham, MA, USA). Similar to previous work with antibiotic-loaded, water-in-PFC emulsions [24 (link),25 (link)], the PFC used was perfluorocycloether/perfluorooctane (FC-770) purchased from 3 M Inc. (St. Paul, MN, USA) and the fluorosurfactant used (“FSH-PEG”) was perfluoropolyether (Krytox 157FSH oil: 7 kDa, n = 41)-polyethelene glycol (PEG: 1 kDa, m = 22)- Krytox 157FSH (i.e., FSH-PEG-FSH) tri-block copolymers synthesized from Krytox 157FS oil purchased from Dupont (Wilmington, DE, USA) [26 (link)]. See Fig. 1 for the molecular structure of the FSH-PEG copolymer.

Nelson D.L., Zhao Y., Fabiilli M.L, & Cook K.E. (2018). In vitro evaluation of lysophosphatidic acid delivery via reverse perfluorocarbon emulsions to enhance alveolar epithelial repair. Colloids and surfaces. B, Biointerfaces, 169, 411-417.

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In Vivo Imaging of T Cell Infiltration in EAE

Cited 3 times

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Th1 or Th17 cells were labeled for 45 min at 37°C with 7-amino-4-chloromethylcoumarin (CMAC) (Molecular Probes). Then, 2 × 10⁷ Th1 or Th17 cells were intravenously injected into MOG_35-55-immunized EAE recipient mice at the onset of the disease. After 48 h, when mice reached the EAE peak, 70 KDa fluorescein isothiocyanate dextran (FITC–dextran) (Sigma Chemical Co., St. Louis, MI, USA) was intravenously injected immediately before imaging of blood vessels. A mode-locked Ti : Sapphire Chameleon Ultra II laser (Coherent Inc.) was tuned to 750–800 nm and an Olympus XLUMPlanFI 20× objective (water immersed, numerical aperture, 0.95) was used for TPLSM imaging. To visualize infiltrating T cells in the dorsal spinal SAS, time-lapse sequences were performed at 70 μm stack height (2.5 μm z step) at 35-40 s intervals as previously described (17 (link)). Immediately after the first acquisition, 200 μl of artificial CSF containing 100 μg/ml of blocking anti-α4 integrins (clone PS/2, rat anti-mouse) or blocking anti-α4β7 integrin (clone DATK32, rat anti-mouse) were locally applied to the exposed spinal cord and incubated for 30 min before a second round of imaging, to allow adequate local meningeal diffusion (17 (link), 23 (link)–26 (link)).

Rossi B., Dusi S., Angelini G., Bani A., Lopez N., Della Bianca V., Pietronigro E.C., Zenaro E., Zocco C, & Constantin G. (2023). Alpha4 beta7 integrin controls Th17 cell trafficking in the spinal cord leptomeninges during experimental autoimmune encephalomyelitis. Frontiers in Immunology, 14, 1071553.

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Tight Junction Protein Localization in MDCK Cells

Cited 2 times

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MDCK II cells were provided by Dr. Masayuki Murata [9 (link)]. MDCK I cells were provided by the late Dr. Shoichiro Tsukita (Kyoto University) and maintained in our laboratory [9 (link)]. Cells were grown in DMEM (high glucose) supplemented with 5% fetal bovine serum.
Mouse anti-ZO-1 monoclonal antibody (mAb) (T8/754), rat anti-occludin mAb (MOC37), rabbit anti-claudin-2 polyclonal antibody (pAb), and rabbit anti-claudin-4 pAb were characterized as described previously [5 (link),22 (link)–24 (link)]. Rabbit anti-ZO-2 pAb (38–9100), rabbit anti-ZO-3 pAb (36–4100), rabbit anti-claudin-1 pAb (51–9000), mouse anti-claudin-2 mAb (32–5600), rabbit anti-claudin-3 pAb (34–1700), mouse anti-claudin-4 mAb (32–9400), rabbit anti-claudin-7 pAb (34–9100), and alexa fluor 488 phalloidin (A12379) were purchased from Invitrogen. Rabbit anti-FLAG pAb (PM020) and rat anti-GFP mAb (D153–3) were purchased from Medical and Biological Laboratories. Mouse anti-FLAG mAb (018–22381) was purchased from Wako. Rabbit anti-nonmuscle myosin heavy chain II-B (MHC-B) pAb (PRB-445P) was purchased from Covance. Mouse anti-E-cadherin mAb (ECCD-2; M108) was purchased from Clontech. Fluorescein isothiocyanate-dextran (FITC-dextran) was purchased from Sigma-Aldrich. Fluorescein (16106–82) was purchased from Nacalai tesque.

Tokuda S, & Furuse M. (2015). Claudin-2 Knockout by TALEN-Mediated Gene Targeting in MDCK Cells: Claudin-2 Independently Determines the Leaky Property of Tight Junctions in MDCK Cells. PLoS ONE, 10(3), e0119869.

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Functionalized Gold Nanoparticle Synthesis

Cited 3 times

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Tetrachloroauric (III) trihydrate (HAuCl₄·3H₂O: 99.9%), sodium citrate tribasic dehydrate (≥99%) and hydroquinone (≥99%) were purchased from Sigma-Aldrich. Heterobifunctional polyethylene glycol OPSS-PEG-SVA (3.4 kDa, OPSS-PEG-SVA-3400) were purchased from Laysan Bio Inc. Poly(ethylene glycol) 2-mercaptoethyl ether acetic acid (Thiol-PEG-Carboxyl, 600 Da, PG2-CATH-600) were purchased from Nanocs Inc. Fluorescein isothiocyanate–dextran (FITC-dextran) and WST-1 cell proliferation reagent (05015944001) was purchased from Sigma-Aldrich. Propidium iodide (PI) is purchased from Cayman chemical (14289). Mouse IgG isotype control (Ctrl Ab, 31901) was purchased from Invitrogen. Anti-PAR2 antibody (MAB3949) was purchased from R&D systems. Anti-JAM-A antibody BV16 was kindly provided by Dr. Elisabetta Dejana’s lab.^{32 (link)} Human cerebral microvessel endothelial cells (hCMEC/D3) were purchased from EMD Millipore. HEK293 cell with Ca²⁺ indicator TN-XXL was kindly provided by Dr. Paul Slesinger’s lab.^{44 (link)} PAR2 agonist 2-aminothiazol-4-yl-LIGRL-NH₂ (2AT) was provided by Dr. Theodore Price’s lab.^{40 (link)} Somatostatin (SST) was purchased from Sigma-Aldrich (S9129). Pierce modified lowery protein assay kit was obtained from ThermoFisher Scientific (23240).

Kang P., Li X., Liu Y., Shiers S.I., Xiong H., Giannotta M., Dejana E., Price T.J., Randrianalisoa J., Nielsen S.O, & Qin Z. (2019). Transient Photo-Inactivation of Cell Membrane Protein Activity without Genetic Modification by Molecular Hyperthermia. ACS nano, 13(11), 12487-12499.

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Dendrimer-based Microscopy Staining Protocols

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The following agents were purchased: hydroxyl terminated ethylenediaminocore PAMAM dendrimer (referred to as dendrimer throughout, unless otherwise specified) (Dendritech, Midland, MI), Methanol (HPLC grade), DMF (HPLC grade), stainless steel beads (Fisher Scientific, Waltham, MA); and Cyanine 5 (Cy5) (GE Healthcare Life Science, Pittsburgh, PA). For confocal microscopy: nuclei counter-stain, 4′,6-diamidino-2-phenylindole, dihydrochloride (DAPI), Alexa Fluor^® 594 Goat Anti-Rabbit IgG (H + L) Antibody (Molecular Probes, Eugene, Oregon); Fluorescent mounting media (Dako, Santa Clara, CA); Anti-Iba1, Rabbit (Wako, Osaka, Japan); Lectin from Bandeiraea simplicifolia (BSI-B4) (Sigma–Aldrich, St. Louis, MO); Anti-GFAP 488 (eBioscience, San Diego, CA); Fluorescein isothiocyanate–dextran (FITCdextran), molecular weight 70,000 Da (Sigma Aldrich, St. Louis, MO).

Zhang F., Mastorakos P., Mishra M.K., Mangraviti A., Hwang L., Zhou J., Hanes J., Brem H., Olivi A., Tyler B, & Kannan R.M. (2015). Uniform brain tumor distribution and tumor associated macrophage targeting of systemically administered dendrimers. Biomaterials, 52, 507-516.

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