T-47D BC cells were grown on coverslips. Cells were fixed for 30 min with paraformaldehyde (4%) and permeabilized for 5 min with 0.1% Triton. Blocking was performed with PBS containing 3% BSA for 30 min. BC cells were incubated with antibodies vs. phospho-FAKY397 and phospho-MoesinT558 (1:500, BD Transduction Laboratories) overnight at 4°C followed by incubation with a fluorescein-conjugated goat anti-rabbit/mouse secondary antibody (1:200; Vector Laboratories). Texas Red-phalloidin (Sigma-Aldrich, St. Louis) was added for 30 min to reveal actin and the nuclei were counterstained with or 4′-6-diamidino-2-phenylindole (Sigma-Aldrich, St. Louis) and mounted with Vectashield mounting medium (Vector Laboratories, Burlingame, CA, USA). Immunofluorescence images were captured using a Nikon Eclipse E200 microscope (Japan) coupled to a high-resolution 590CU 5.0M CCD digital camera.
Fluorescein conjugated goat anti rabbit mouse secondary antibody
Manufactured by Vector Laboratories
Sourced in Canada
Fluorescein-conjugated goat anti-rabbit/mouse secondary antibody is a laboratory reagent used for the detection of rabbit or mouse primary antibodies in various immunoassay techniques. It contains goat-derived antibodies that are conjugated with the fluorescent dye fluorescein, enabling the visualization of target antigens.
Automatically generated - may contain errors
2 protocols using fluorescein conjugated goat anti rabbit mouse secondary antibody
1
Immunofluorescence Imaging of Phosphorylated FAK and Moesin in T-47D Breast Cancer Cells
2
Immunofluorescence Staining of T-47D Cells
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T-47D cells were grown on coverslips. Cells were fixed with 4% paraformaldehyde for 30 min and permeabilized with 0.1% Triton for 5 min. Blocking was performed with PBS containing 3% bovine serum albumin for 30 min. Cells were incubated with antibodies against Tyr 397 -phospho-FAK (Transduction Laboratories, Lexington, KY), Vinculin (sc-7648), and Thr 558 -p-Moesin (sc-12895) (Santa Cruz Biotechnology, Santa Cruz, CA) overnight at 4º C followed by incubation with a fluorescein-conjugated goat anti-rabbit/mouse secondary antibody (1:200; Vector Laboratories). Then cells were incubated with Texas Red-phalloidin to stain actin fibers (Sigma) for 30 min. After washing, the nuclei were counterstained with
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