Total splenocytes and Min-6 cells were washed (with PBS supplemented with 0.5% BSA) and incubated with fluorochrome labeled anti CD4-efluor 450, anti-CTLA-4 PE (ebioscience, San Diego, CA); fixed and permeabilized cells were incubated with unlabelled guinea pig-derived anti-insulin antibody (Zymed Laboratories, South San Francisco, CA), purified anti-GLUT2 (H5, E7 and D2 mAbs), anti-CTLA-4 mAbs and BsAbs in different combinations on ice for 30 min. Some samples were further washed and probed with fluorochrome labeled anti-guinea pig IgG-TRITC (Zymed Laboratories, South San Francisco, CA), anti-mouse IgG-FITC or anti-Hamster. FITC-IgG (ebioscience, San Diego, CA) stained cells were washed three times and analyzed by Cyan flow cytometer (Beckman/Coulter). For analysis of intracellular Foxp3, cells were fixed/permeabilized and incubated with anti-Foxp3-APC antibodies.
Fitc igg
Manufactured by Thermo Fisher Scientific
Sourced in United States
FITC-IgG is a fluorescently labeled immunoglobulin (IgG) conjugate. FITC, which stands for Fluorescein Isothiocyanate, is the fluorescent dye attached to the IgG molecule. This product can be used as a detection reagent in various immunoassay and cell biology applications.
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Lab products found in correlation
3 protocols using fitc igg
1
Multiparametric Immunophenotyping of Islet Cells
2
Evaluating Monocytic Differentiation by Flow Cytometry
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Cell differentiation was detected using the APC-labeled anti-human CD14 (#11-0118-42; eBioscience, San Diego, CA, USA) and FITC or PE-labeled anti-human CD11b (#17-0149-42 or 12-0112-82, eBioscience) antibodies or the corresponding isotype control antibody (FITC-IgG, #11-4714-41, eBioscience; or APC-IgG, #400119, BioLegend, San Diego, CA, USA) and analyzed by FCM.
Cells were plated in 6-well plates at a density of 5 × 105 cells/well and treated for 5 days with either DMSO (0.1%) or HEL (1, 5, or 10 μmol/L). ATRA (1 μmol/L) was used as a positive control. The cell medium was replaced with fresh medium after 3 days of treatment.
For the FCM differentiation assay, cells were washed with ice-cold flow cytometry staining buffer (FCS, #00-4222-26, eBioscience) twice, resuspended in 100 μL FCS buffer, and then stained with FITC-anti-human CD11b or APC-anti-human CD14 for 45 min on ice. Appropriate isotype controls were included in this analysis. After incubation, cells were washed twice with FCS buffer, resuspended in 0.3 mL FCS buffer, and analyzed by FCM using the Gallios Flow Cytometer. FlowJo software was used to analyze the results. The sample size was n = 3 for each treatment condition.
Cells were plated in 6-well plates at a density of 5 × 105 cells/well and treated for 5 days with either DMSO (0.1%) or HEL (1, 5, or 10 μmol/L). ATRA (1 μmol/L) was used as a positive control. The cell medium was replaced with fresh medium after 3 days of treatment.
For the FCM differentiation assay, cells were washed with ice-cold flow cytometry staining buffer (FCS, #00-4222-26, eBioscience) twice, resuspended in 100 μL FCS buffer, and then stained with FITC-anti-human CD11b or APC-anti-human CD14 for 45 min on ice. Appropriate isotype controls were included in this analysis. After incubation, cells were washed twice with FCS buffer, resuspended in 0.3 mL FCS buffer, and analyzed by FCM using the Gallios Flow Cytometer. FlowJo software was used to analyze the results. The sample size was n = 3 for each treatment condition.
3
Fluorescent Staining and Flow Cytometry Analysis of Lymphocytes
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Single-cell preparations of spleens and mesenteric lymph nodes were prepared from tissues recovered from sacrificed animals in experiment 2 [31 (link)]. Approximately 106 cells were surface stained with fluorescein isothiocyanate-labeled goat anti-Syrian hamster immunoglobulin G (FITC-IgG) and phycoerythrin-labeled anti-mouse CD4 [PE-L3T4; eBiosciences (San Diego, CA)] previously shown to interact with hamster cells [31 (link),34 (link)]. Cell surface determinant data were acquired for 105 cells per sample, using a FACSCalibur flow cytometer (BD Biosciences, San Jose, CA). Lymphocytes were selected by forward scatter gating on size, and data were analyzed using the FlowJo software program (Treestar, Ashland, OR).
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