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Fluorescein isothiocyanate (fitc)

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FITC is a fluorescent dye commonly used in biological research. It is a derivative of fluorescein, which emits a green fluorescent signal when excited by light of a specific wavelength. FITC can be conjugated to a variety of biomolecules, such as antibodies, proteins, and nucleic acids, to enable their detection and visualization in various experimental applications.

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Lab products found in correlation

Donkey anti chicken dylight 649, by Jackson ImmunoResearch (3 mentions) Accuri c6 flow cytometer, by BD (3 mentions) Goat serum, by Thermo Fisher Scientific (1 mentions) Texas red, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Phycoerythrin pe, by Thermo Fisher Scientific (1 mentions) Facscalibur, by BD (1 mentions) Cellquest software, by BD (1 mentions)

Close spelling variants of this product

Peanut agglutinin conjugated to fluorescein-isothiocyanate (PNA-FITC) (2 citations) Phalloidin-fluorescein isothiocyanate (FITC) (2 citations)

6 protocols using fluorescein isothiocyanate (fitc)

1

Quantifying αGal and Neu5Gc Levels

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Example 18

Isolectin B4 (IB4) counter-selected LDC, cloned fetal fibroblasts from DKO fetuses, wild-type LDC and wild-type fetal fibroblasts were stained with IB4 conjugated with FITC (Enzo Life Science, Farmingdale N.Y.) to assess the αGal epitope level. To evaluate the Neu5Gc level, IB4 counter-selected LDC cloned fetal fibroblasts from DKO fetuses, wild-type LDC and wild-type fetal fibroblasts were stained with antiNeu5Gc antibody (Sialix, Vista Calif.) followed by donkey anti-chicken DyLight 649 (Jackson ImmunoResearch Laboratories Inc., West Grove Pa.). A negative control antibody for comparison with anti-Neu5Gc antibody was also used (Sialix, Vista Calif.). An Accuri C6 flow cytometer (Accuri, Ann Arbor Mich.) and FlowJo software (Tree Star, Inc. Ashland Oreg.) were used for analysis. Representative flow cytometry results of αGal from IB4 counter selected cells are shown in FIG. 7A. Representative flow cytometry results of Neu5Gc on IB4 counter selected cells are shown in FIG. 8A. Representative flow cytometry results of the αGal and Neu5Gc on fetal fibroblasts derived from fetus 7 are shown in FIG. 9C.

US11666039B2. Double knockout (GT/CMAH-KO) pigs, organs and tissues (2023-06-06). Indiana University Research and Technology Corporation [US]. Inventors: A. Joseph Tector [US].
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2

Quantifying αGal and Neu5Gc Epitopes in Fetal Fibroblasts

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Example 18

Isolectin B4 (IB4) counter-selected LDC, cloned fetal fibroblasts from DKO fetuses, wild-type LDC and wild-type fetal fibroblasts were stained with IB4 conjugated with FITC (Enzo Life Science, Farmingdale N.Y.) to assess the αGal epitope level. To evaluate the Neu5Gc level, IB4 counter-selected LDC cloned fetal fibroblasts from DKO fetuses, wild-type LDC and wild-type fetal fibroblasts were stained with antiNeu5Gc antibody (Sialix, Vista Calif.) followed by donkey anti-chicken DyLight 649 (Jackson ImmunoResearch Laboratories Inc., West Grove Pa.). A negative control antibody for comparison with anti-Neu5Gc antibody was also used (Sialix, Vista Calif.). An Accuri C6 flow cytometer (Accuri, Ann Arbor Mich.) and FlowJo software (Tree Star, Inc. Ashland Oreg.) were used for analysis. Representative flow cytometry results of αGal from IB4 counter selected cells are shown in FIG. 7A. Representative flow cytometry results of Neu5Gc on IB4 counter selected cells are shown in FIG. 8A. Representative flow cytometry results of the αGal and Neu5Gc on fetal fibroblasts derived from fetus 7 are shown in FIG. 9C.

US10667500B2. Double knockout (GT/CMAH-KO) pigs, organs and tissues (2020-06-02). Indiana University Research and Technology Corporation [US]. Inventors: A. Joseph Tector [US].
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3

Nuclear Translocation of NRF2 and KEAP1

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The nuclear translocation of NRF2 and KEAP1 was detected via immunofluorescence staining. PBL cells (1 × 104/ml) were cultured in laser confocal Petri dishes, treated with andrographolide sodium bisulfate (ASB) for 1 d, and irradiated with UV (300 μW/cm2 sec x 300 sec). PBL were washed 4 times with PBS and fixed with 4% paraformaldehyde for 20 min at room temperature, permeabilized with 1% Triton X for 20 min, and then washed and blocked with 5% goat serum (Invitrogen, Frederick, MD, USA) for half an hour. PBL were incubated with primary antibodies (NRF2, 1 : 2000, and KEAP1, 1 : 2000) at 4°C overnight, washed with TBST from G-Biosciences (St. Louis, MO, USA), and incubated with a secondary antibody conjugated to a fluorochrome (FITC, Enzo Life Sciences, NY, USA) for 2 h at room temperature. PBL were stained with DAPI (10 μg/ml) for 10 min, washed with PBST 3 times, and drawn on the slides with a drop of the fluorescent mounting medium. The antibody localization was visualized using a fluorescence microscope (EVOS; Life Technologies, USA).
Wen C., Jiang M., Huang W, & Liu S. (2020). Antarctic Krill Oil Attenuates Oxidative Stress via the KEAP1-NRF2 Signaling in Patients with Coronary Heart Disease. Evidence-based Complementary and Alternative Medicine : eCAM, 2020, 9534137.
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4

Flow Cytometry Analysis of Gal and Neu5Gc Levels

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Example 18

Isolectin B4 (IB4) counter-selected LDC, cloned fetal fibroblasts from DKO fetuses, wild-type LDC and wild-type fetal fibroblasts were stained with IB4 conjugated with FITC (Enzo Life Science, Farmingdale N.Y.) to assess the αGal epitope level. To evaluate the Neu5Gc level, IB4 counter-selected LDC cloned fetal fibroblasts from DKO fetuses, wild-type LDC and wild-type fetal fibroblasts were stained with antiNeu5Gc antibody (Sialix, Vista Calif.) followed by donkey anti-chicken DyLight 649 (Jackson ImmunoResearch Laboratories Inc., West Grove Pa.). A negative control antibody for comparison with anti-Neu5Gc antibody was also used (Sialix, Vista Calif.). An Accuri C6 flow cytometer (Accuri, Ann Arbor Mich.) and FlowJo software (Tree Star, Inc. Ashland Oreg.) were used for analysis. Representative flow cytometry results of αGal from IB4 counter selected cells are shown in FIG. 7A. Representative flow cytometry results of Neu5Gc on IB4 counter selected cells are shown in FIG. 8A. Representative flow cytometry results of the αGal and Neu5Gc on fetal fibroblasts derived from fetus 7 are shown in FIG. 9C.

US09888674B2. Double knockout (GT/CMAH-KO) pigs, organs and tissues (2018-02-13). None [US]. Inventors: A. Joseph Tector [US].
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5

Multicolor Fluorescent Labeling of BAC Clones

Cited 1 time
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BAC clones labeling was carried out with a first amplification by DOP-PCR, followed by a conventional PCR for labeling, as described previously in Garcia Angulo et al.28 (link). Three different fluorochromes were used: Texas red (Thermo Fisher Scientific, USA), fluorescein-isothiocyanate (FITC) (Enzo, USA), and diethyl-aminocoumarin (DEAC) (Vysis, USA). The chromosomes were pretreated with pepsin and fixed in formaldehyde. Finally, the chromosome preparation was dehydrated with ethanol series and air-dried before hybridization. Hybridization was done according to Portela-Bens et al.29 (link).
García E., Cross I., Portela-Bens S., Rodríguez M.E., García-Angulo A., Molina B., Cuadrado A., Liehr T, & Rebordinos L. (2019). Integrative genetic map of repetitive DNA in the sole Solea senegalensis genome shows a Rex transposon located in a proto-sex chromosome. Scientific Reports, 9, 17146.
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6

Flow Cytometric Analysis of DC Activation

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DCs were treated with 50 µg/ml of the ethanol-killed S. aureus strain for 48 h, or with Pam3CSK4 at 0–10 μg/ml for 0 to 72 h. Following stimulation, the cells were stained with anti-mouse antibodies conjugated with fluorescent dyes as previously described (33 (link)). For membrane-bound BAFF (mBAFF) analysis, DCs were stained with anti-mouse BAFF antibodies conjugated with fluorescein isothiocyanate (FITC) (Enzo Life Science, Plymouth Meeting, PA, USA) together with anti-mouse CD11c antibodies conjugated with allophycocyanin (APC) (BD Biosciences). Additionally, DCs were stained with antibodies for mouse TLR1 conjugated with Alexa Fluor 647 or TLR2 conjugated with phycoerythrin (PE) (e-Bioscience, San Diego, CA, USA). The cells were then subjected to flow cytometry (FACSCalibur) with CellQuest software (BD Biosciences). Data were analyzed using FlowJo software (Tree Star, San Carlos, CA, USA).
Im J., Baik J.E., Lee D., Park O.J., Park D.H., Yun C.H, & Han S.H. (2020). Bacterial Lipoproteins Induce BAFF Production via TLR2/MyD88/JNK Signaling Pathways in Dendritic Cells. Frontiers in Immunology, 11, 564699.
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