Cells at a density of 1 × 105 were briefly plated in 6-well plates, and three glass coverslips were placed onto the wells. The plates were incubated for 24–48 h until 30–40% confluence was reached. The cells were then fixed with 4% paraformaldehyde for 10 min at room temperature and permeabilized with 0.1% Triton X-100. The slides were then washed three times with phosphate-buffered saline (PBS) and blocked with 5% bovine serum albumin in PBS for 30 min at room temperature. The sections were incubated with a primary antibody against PLAC8 overnight. After rinsing three times with PBST for 5 min, an Alexa 488-conjugated (green) goat anti-rabbit antibody was applied (1:200 dilution; Life Technologies) for 1 h at room temperature. Cell nuclei were counterstained with DAPI (4',6-diamidino-2-phenylindole) and images were acquired using a Nikon laser scanning confocal microscope (Nikon Instruments Inc., Melville, NY, USA).
Alexa 488 conjugated green goat anti rabbit antibody
Manufactured by Thermo Fisher Scientific
Alexa Fluor 488-conjugated goat anti-rabbit antibody is a secondary antibody used for immunodetection applications. It is designed to bind to and detect rabbit primary antibodies, producing a green fluorescent signal.
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2 protocols using alexa 488 conjugated green goat anti rabbit antibody
1
Immunofluorescence Analysis of PLAC8 Protein
2
Immunofluorescence Staining of PLAC8
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Cells at a density of 1 × 10 5 were brie y plated in 6-well plates, and three glass coverslips were placed onto the wells. The plates were incubated for 24-48 h until 30-40% con uence was reached. The cells were then xed with 4% paraformaldehyde for 10 min at room temperature and permeabilized with 0.1% Triton X-100. The slides were then washed three times with phosphate-buffered saline (PBS) and blocked with 5% bovine serum albumin in PBS for 30 min at room temperature. The sections were incubated with a primary antibody against PLAC8 overnight. After rinsing three times with PBST for 5 min, an Alexa 488conjugated (green) goat anti-rabbit antibody was applied (1:200 dilution; Life Technologies) for 1 h at room temperature. Cell nuclei were counterstained with DAPI (4',6-diamidino-2-phenylindole), and images were acquired using a Nikon laser scanning confocal microscope (Nikon Instruments Inc., Melville, NY, USA).
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