MSCs were fixed in 4% PFA at room temperature for 15 min and permeabilized with 0.5% Triton X-100 for 5 min. The cells were blocked with 2.5% goat serum for 1 h and then stained with 1:200 anti-Ki-67 mouse monoclonal antibody (M7240; Dako; Agilent Technologies, Inc.) overnight at 4°C. Antibody binding was visualized by incubation with a FITC-conjugated goat anti-mouse secondary antibody (1:200, sc-2010; Santa Cruz Biotechnology, Inc.) at room temperature for 1 h. Mouse IgG2a (1:200, sc-2856; Santa Cruz Biotechnology, Inc.) was used as the isotype control for Ki-67 antibody and FITC-conjugated goat anti-mouse antibody (1:200, sc-2010; Santa Cruz Biotechnology, Inc.) as a negative control to assess non-specific binding of the secondary antibody. MSCs were then stained with DAPI at room temperature for 3 min and imaged using an Olympus BX51 fluorescence microscope.
Fitc conjugated goat anti mouse secondary antibody
Manufactured by Santa Cruz Biotechnology
FITC-conjugated goat anti-mouse secondary antibody is a laboratory tool used for the detection of mouse primary antibodies. It functions by binding to the Fc region of mouse antibodies and emitting a fluorescent signal upon excitation, enabling the visualization and identification of target proteins or molecules in various research applications.
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Lab products found in correlation
Bx51 fluorescence microscope, by Olympus (1 mentions)
Mouse igg2a, by Santa Cruz Biotechnology (1 mentions)
Fitc conjugated goat anti mouse antibody, by Santa Cruz Biotechnology (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
2 protocols using fitc conjugated goat anti mouse secondary antibody
1
Immunofluorescent Staining of MSCs
2
Immunofluorescence Staining of SH3BGRL
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Cells were fixed with 4% paraformaldehyde and then permeabilized with 0.5% Triton. Samples were blocked with 1% BSA for 30 min at room temperature and stained with mouse anti-SH3BGRL monoclonal antibody overnight at 4°C, followed by incubation for 1 h at room temperature with fluorescein (FITC)-conjugated goat antimouse secondary antibody (Santa Cruz Biotechnology) in the dark. Finally, the samples were mounted with an antifade reagent with DAPI (Invitrogen, Carlsbad, CA). Imaging was conducted with a fluorescence microscope (Nikon, Japan).
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