HeLa cells were plated on coverslips and harvested at 48 h posttransfection. After being washed twice with phosphate-buffered saline (PBS), cells were fixed with 4% paraformaldehyde diluted by PBS and permeabilized using 0.1% Triton X-100. Then fixed cells were washed five times with PBS and incubated in blocking solution for 2 h. After washing off the blocking solution, cells were incubated with anti-HA (1:200; cat. no. H6908, Sigma) and anti-Flag (1:100; cat. no. F1804, Sigma) diluted in PBS containing 0.1% bovine serum albumin (BSA) for 2 h. Cells were washed three times with PBST and incubated with fluorescein (FITC) goat anti-mouse IgG (1:500; cat. no. 115-095-003, Jackson ImmunoResearch) and rhodamine (TRITC) goat anti-rabbit IgG (1:500; cat. no. 111-025-003, Jackson ImmunoResearch) for 1 h. Following four washes with PBS, cells were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, 1:2,000), and mounted on slides with Antifade Mounting Medium (Beyotime) for confocal microscopy analysis. The images were captured by Leica TCS SP5 confocal system.
Fluorescein fitc goat anti mouse igg
Manufactured by Merck Group
Fluorescein (FITC) goat anti-mouse IgG is a secondary antibody conjugated with the fluorescent dye fluorescein isothiocyanate (FITC). It is designed to detect and visualize mouse immunoglobulin G (IgG) antibodies in various immunoassays and microscopy applications.
Automatically generated - may contain errors
2 protocols using fluorescein fitc goat anti mouse igg
1
Immunofluorescence Staining of Transfected HeLa Cells
2
Immunofluorescence Staining of Transfected HeLa Cells
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HeLa cells were plated on coverslips and harvested at 48 h posttransfection. After being washed twice with phosphate-buffered saline (PBS), cells were fixed with 4% paraformaldehyde diluted by PBS and permeabilized using 0.1% Triton X-100. Then fixed cells were washed five times with PBS and incubated in blocking solution for 2 h. After washing off the blocking solution, cells were incubated with anti-HA (1:200; cat. no. H6908, Sigma) and anti-Flag (1:100; cat. no. F1804, Sigma) diluted in PBS containing 0.1% bovine serum albumin (BSA) for 2 h. Cells were washed three times with PBST and incubated with fluorescein (FITC) goat anti-mouse IgG (1:500; cat. no. 115-095-003, Jackson ImmunoResearch) and rhodamine (TRITC) goat anti-rabbit IgG (1:500; cat. no. 111-025-003, Jackson ImmunoResearch) for 1 h. Following four washes with PBS, cells were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, 1:2,000), and mounted on slides with Antifade Mounting Medium (Beyotime) for confocal microscopy analysis. The images were captured by Leica TCS SP5 confocal system.
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