Differentiated RGC cells were used for phenotypic marker identification by flow cytometry. Trypsinized cells were resuspended in 100 μL of PBS and incubated with primary antibodies (anti-human CD90) (1:100 dilutions) at 4 °C for 1 h. After washing twice with PBS, labeled cells were resuspended in 100 μL of PBS with 1 μL of the fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG antibody (Millipore) at 4 °C for 1 h. Cells were then analyzed using a BD FACSCalibur apparatus (BD Biosciences, San Jose, CA, USA).
Fitc conjugated goat anti mouse igg antibody
Manufactured by Merck Group
Sourced in United States
FITC-conjugated goat anti-mouse IgG antibody is a laboratory reagent used for the detection and quantification of mouse immunoglobulin G (IgG) in various immunoassay applications. The antibody is conjugated with the fluorescent dye fluorescein isothiocyanate (FITC), which allows for the visualization and analysis of the target mouse IgG.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 700, by Zeiss (2 mentions)
Facscalibur apparatus, by BD (1 mentions)
Pezgs0416, by Merck Group (1 mentions)
Rabbit anti human fxr antibody, by Abcam (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Te2000 e, by Nikon (1 mentions)
Oct compound, by Sakura Finetek (1 mentions)
5 protocols using fitc conjugated goat anti mouse igg antibody
1
Phenotypic Profiling of Differentiated RGCs
2
Immunofluorescent Staining of FXR
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Cells were seeded in 4-well chamber slides (#PEZGS0416, Millipore, USA), washed twice with cold PBS, fixed with 500 μl 4% paraformaldehyde for 35 min, permeabilized with 200 μl 1% Triton X-100 for 10 min, and blocked with 500 μl goat serum for 30 min. Next, the cells were incubated with primary antibodies at 4 °C overnight. Subsequently, the cells were incubated with FITC-conjugated goat anti-mouse IgG antibody (#12–506, 1:200, Millipore, USA) at room temperature for 2 h. Finally, the nuclei were stained with DAPI (#C0060, 1:800, Solarbio, China) for 10 min. The primary antibody used was a rabbit anti-human FXR antibody (#187735, 1:100, Abcam, UK).
3
Immunofluorescence Staining of c-Myc Protein
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MDCK cells were washed 3 times with PBS and fixed in cold fixative solution (acetone: methanol, 3:1) for 10 min at 4°C followed by 3 washes with PBS and incubation for 1 h at 37°C with individual primary antibodies. The cells were washed 3 times and incubated with purified primary mouse anti-c-Myc Epitope Tag mAb (MBL: 1:1000 dilution) as the second antibody. The cells were again washed 3 times, incubated with fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG antibody (Sigma, 1:100 dilution) for 1 h at 37°C, washed a further 3 times with PBS, and stained with 100 μg/ml propidium iodide (PI) for 5 min. Fluorescence was observed with a confocal laser scanning microscope (Nikon TE2000-E).
4
Renal Immune Complex Deposition Analysis
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The kidneys isolated from the host mice were embedded in OCT compound (Sakura Finetek, Alphen aan den Rijn, Netherlands) and snap-frozen on dry ice; sections (8 µm) were prepared on glass slides, fixed in acetone and ethanol for 5 min, and dried. After blocking with PBS containing 10% fetal calf serum for 30 min at room temperature, the sections were incubated with FITC-conjugated goat anti-mouse C3 antibody (MP Biomedicals, Irvine, CA, USA) diluted 1:500 in FACS buffer for 2 h at room temperature in the dark. To detect IC deposition, the sections were incubated with FITC-conjugated goat anti-mouse IgG antibody (Sigma Aldrich, St. Louis, MO, USA) diluted 1:320 in FACS buffer for 2 h at room temperature. The nucleus was stained using 4′,6-diamidino-2-phenylindole. Fluorescent staining was observed using a fluorescent microscope (Zeiss LSM700, Oberkochen, Germany). Fluorescence intensity was evaluated using ImageJ software (version 1.52a, National Institutes of Health, Bethesda, MD, USA).
5
Detecting Avian Leukosis Virus Infection
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Cells infected with or without virus were fixed with an acetone:ethanol solution (3:2) for 10 min after 7 days and washed once with PBS. After blocking with 1% BSA in PBS, the fixed cells were incubated with the mouse anti-ALV p27 mAb 5D3 for 45 min at 37 °C. After three washes with PBS, the cells were incubated with a FITC-conjugated goat anti-mouse IgG antibody (Sigma-Aldrich, USA) for another 45 min. After three washes with PBS, the cells were observed under a fluorescence microscope.
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