For co-localization analysis, HCT116 cells were plated on sterilized coverslips, rinsed with PBS and fixed with 4% paraformaldehyde for 15 min at room temperature. Then the cells were permeabilized with 0.25% Triton X-100 in PBS at room temperature for 10 min. After an incubation with blocking buffer (0.5% bovine serum albumin in PBS) for 30 min, the coverslips were simultaneously co-incubated with two primary antibodies: a mouse monoclonal antibody against MTA1 (Abcam) and a rabbit polyclonal antibody against Ku70 (Santa Cruz) or Ku80 (Santa Cruz, USA) over night at 4 °C. After washing with PBS, cells were then co-incubated with a FITC-conjugated goat anti-mouse antibody (ZSGB-BIO) and a TRITC-conjugated goat anti-rabbit antibody (ZSGB-BIO) for 1 h at room temperature. The coverslips were then mounted with mounting medium containing DAPI (ZSGB-BIO). And the fluorescent images were acquired using a fluorescent microscope (Leica).
Fitc conjugated goat anti mouse antibody
Manufactured by ZSGB-BIO
FITC-conjugated goat anti-mouse antibody is a laboratory reagent used for the detection and identification of mouse-derived proteins or other biomolecules in various experimental techniques. It consists of a goat-derived antibody that specifically binds to mouse immunoglobulins, conjugated with the fluorescent dye FITC (Fluorescein Isothiocyanate) to enable visualization and signal detection.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescent microscope, by Leica (1 mentions)
Mouse monoclonal antibody against mta1, by Abcam (1 mentions)
Rabbit polyclonal antibody against ku70, by Santa Cruz Biotechnology (1 mentions)
Anti p47phox antibody, by Santa Cruz Biotechnology (1 mentions)
Confocal laser scanning microscopy, by Zeiss (1 mentions)
2 protocols using fitc conjugated goat anti mouse antibody
1
Co-localization Analysis of MTA1 and Ku Proteins
2
Immunocytofluorescence Microscopy for Protein Localization
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Immunocytofluorescence was performed as in our previous study, with some modifications (Tong et al., 2017 (link)). Culture supernatant was discarded, and cells were then washed three times with pre-chilled PBS. Cells were fixed in cold 4% paraformaldehyde in PBS for 20 min and then washed three times with cold PBS. Cell slides were blocked with 5% BSA containing 0.3% Triton X-100 in PBS for 30 min at room temperature. Cells were incubated with primary antibodies at 4°C overnight as follows: anti-p47 phox antibody (1:100; Santa Cruz Biotechnology Inc.) and anti-nuclear factor-κB (NF-κB) antibody (1:100; Santa Cruz Biotechnology Inc.). The slides were incubated with FITC-conjugated goat anti-mouse antibody (1:300; ZSGB-BIO), and then kept in a dark place at room temperature. Images were acquired using laser scanning confocal microscopy (Zeiss), with the same settings for all samples in each experiment.
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