Fitc isomer 1

Manufactured by Thermo Fisher Scientific

Sourced in United States

FITC isomer I is a fluorescent dye commonly used in biological research. It is a green-fluorescent isomer of fluorescein isothiocyanate (FITC). FITC isomer I can be used for various labeling and detection applications in cell biology, flow cytometry, and fluorescence microscopy.

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Lab products found in correlation

Facscalibur, by BD (1 mentions) Ficoll paque plus, by Cytiva (1 mentions) Accuri flow cytometer, by BD (1 mentions) Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions) Triton x 100, by Merck Group (1 mentions) Sodium deoxycholate, by Merck Group (1 mentions) Ultrapure water system, by Merck Group (1 mentions) C acrylamide bis acrylamide, by Merck Group (1 mentions) Triton x 100 bp 151, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Fluorescein isothiocyanate isomer I (FITC, (2 citations) Fluorescein-5-Isothiocyanate Isomer I (FITC) (2 citations)

4 protocols using fitc isomer 1

Phagocytosis of Staphylococcus by PMNs

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Overnight-grown staphylococci were washed, sonicated, fixed, and fluorescein isothiocyanate (FITC isomer I; Invitrogen)-labeled as described before [79 (link)]. Sample preparation and detection of internalized staphylococci by EA.hy 926 cells were performed by flow-cytometric internalization assays as described before [79 (link)].
The phagocytosis assay was performed as described [87 (link)]. Briefly, PMNs were freshly isolated from Na citrate-treated blood from healthy donors by density gradient centrifugation using Ficoll-Paque Plus (Amersham Bioscience) according to the manufacturer's instruction. FITC-labeled bacteria were added to the PMNs and incubated. Samples were analyzed on a FacsCALIBUR (BD Bioscience). Electronic gating was used to analyze 5,000 PMNs in each sample. The FL1 photomultiplier (transmittance at 500 nm) was used to detect uptake of staphylococcal cells by PMNs.

Bleiziffer I., Eikmeier J., Pohlentz G., McAulay K., Xia G., Hussain M., Peschel A., Foster S., Peters G, & Heilmann C. (2017). The Plasmin-Sensitive Protein Pls in Methicillin-Resistant Staphylococcus aureus (MRSA) Is a Glycoprotein. PLoS Pathogens, 13(1), e1006110.

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Isolation and Labeling of Photoreceptor Outer Segments

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Dissected and homogenized bovine neuroretinas were placed in 20% sucrose and loaded onto a discontinuous 20–60% sucrose gradient. The gradient was ultracentrifuged at 75,600× g for 1 h at 10 °C and the photoreceptor outer segments (POS) collected from the 40% layer. The POS were washed twice with HBSS (ThermoFischer Scientific), resuspended in 2.5% sucrose and the concentration determined by flow cytometry (BD Accuri flow cytometer, BD Biosciences). For the fluorescent labeling, the POS were washed in 0.1 M NaHCO₃, pH = 9, centrifuged at 21,130× g for 10 min at RT and resuspended in 0.1 M NaHCO₃ containing 1:10 dilution of FITC Isomer I (Invitrogen). The samples were incubated overnight at 4 °C in the dark, then washed in PBS and resuspended in ES media. For the phagocytosis assay, two 0.32 cm² wells of iPSC-derived RPE per condition were incubated with 10 POS/cell for 2.5 h at 37 °C, washed, dissociated and analyzed by flow cytometry.

Erkilic N., Gatinois V., Torriano S., Bouret P., Sanjurjo-Soriano C., De Luca V., Damodar K., Cereso N., Puechberty J., Sanchez-Alcudia R., Hamel C.P., Ayuso C., Meunier I., Pellestor F, & Kalatzis V. (2019). A Novel Chromosomal Translocation Identified due to Complex Genetic Instability in iPSC Generated for Choroideremia. Cells, 8(9), 1068.

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Phagocytosis of Photoreceptor Outer Segments by hiPSC-RPE

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MERTK plays an important role in phagocytosis of photoreceptor outer segments (Lukovic et al., 2015 (link)). Phagocytosis in hiPSC-RPE was studied using a slightly modified version of the assay previously described (Mao & Finnemann, 2013 (link)). Bovine retinal photoreceptor rod outer segments (POS) (InVision Bioresources, Seattle, USA) were labeled with FITC isomer I (Invitrogen Corporation, Carlsbad, CA) (Singh et al., 2013 (link)). FITC-labeled POS were seeded onto the mature hiPSC-RPE monolayers grown on Matrigel™ Matrix at 1×10⁷ POS/ml and incubated. In order to detect the internalized POS, the cells were stained with trypan blue, washed with phosphate buffer saline (PBS) and fixed in paraformaldehyde (PFA). Subsequently, the cells were stained with DAPI and Phalloidin; images were captured using Nikon A1RStorm confocal microscope (Nikon, Tokyo, Japan). The number of intracellular FITC labeled POS in hiPSC-RPE were counted using ImageJ 1.52a software. The difference between patient and control groups was evaluated with student’s t test.

Biswas P., Borooah S., Matsui H., Voronchikhina M., Zhou J., Zawaydeh Q., Raghavendra P.B., Ferreyra H., Riazuddin S.A., Wahlin K., Frazer K.A, & Ayyagari R. (2020). Detection and validation of novel mutations in MERTK in a simplex case of retinal degeneration using WGS and hiPSC-RPEs model. Human mutation, 42(2), 189-199.

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Polyacrylamide Gel Electrophoresis Reagents

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2,2-Azobis[2-methyl-N-(2-hydroxyethyl) pro-pionamide] (VA-086, 1% (w/v), photoinitiator) was purchased from Waco Chemical (Richmond, VA). Tetramethy-lethylenediamine (TEMED, T9281), ammonium persulfate (APS, A3678), β-mercaptoethanol (M3148), and a polymer precursor solution composed of 30% T and 2.7% C acrylamide/bis-acrylamide (37.5:1, A3699) were purchased from Sigma-Aldrich (St. Louis, MO). Triton X-100 (BP-151) was purchased from Thermo Fisher Scientific (Waltham, MA). Premixed 10× Tris/glycine/SDS-PAGE buffer (25 mM Tris, pH 8.3; 192 mM glycine; 0.1% SDS) was purchased from BioRad (Hercules, CA). Deionized water (18.2 MΩ) was obtained using an Ultrapure water system (Millipore, Burlington, MA). The cell lysis and PAGE buffer contained 0.5% SDS, 0.1% (v/v) Triton X-100, and 0.25% sodium deoxycholate (D6750, Sigma-Aldrich) in 12.5 mM Tris and 96 mM glycine (pH 8.3, 0.5× from a 10× stock; 161-0734, Bio-Rad, Hercules, CA). FITC (Isomer-I) used for polyacrylamide-gel imaging was from Invitrogen (Carlsbad, CA).

Pan Q., Yamauchi K.A, & Herr A.E. (2018). Controlling Dispersion during Single-Cell Polyacrylamide-Gel Electrophoresis in Open Microfluidic Devices. Analytical chemistry, 90(22), 13419-13426.

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