A549 cells were seeded on sterile confocal petri dish (Biosharp), fixed in methyl alcohol for 15 min, permeabilized by 0.1% Triton X-100, and then blocked with 1% BSA for 30 min. The cells were then incubated with rabbit monoclonal to HuR (Abcam) overnight at 4 °C. After washing three times with PBST, the cells were probed with Alexa-Fluor-555-conjugated goat anti-rabbit-IgG (Abcam) for 1 h at room temperature, followed by nuclear counterstaining with DAPI (Beyotime). Coverslips were observed with a fluorescence microscope (ZEISS, Germany).
Alexa fluor 555 conjugated goat anti rabbit igg
Manufactured by Abcam
Alexa Fluor 555-conjugated goat anti-rabbit IgG is a secondary antibody that specifically binds to rabbit primary antibodies. It is labeled with the Alexa Fluor 555 fluorescent dye, which can be detected using appropriate instrumentation.
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2 protocols using alexa fluor 555 conjugated goat anti rabbit igg
1
Immunocytochemistry of HuR in A549 cells
2
Phospho-Histone H3 Staining in wing discs
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Ten wing discs of rok1167/TM6B and rok1167/167 from third-instar larvae were dissected in the cold PBS buffer. Then, tissues were fixed with a mixture of 3.7% formaldehyde diluted in PBS for 15 min. Heptane was subsequently added for 30 min at room temperature. After washing three times with 1 X PBST (0.1% Triton X-100), a rabbit anti-phospho-Histone H3 (1:200, Sigma, Saint Louis, USA) antibody was incubated overnight at 4 °C. After washing three times with 1 X PBST (0.1% Triton X-100), Alexa Fluor 555 conjugated goat antirabbit IgG (1:200, Abcam) and DAPI were incubated for 1 h. Fluorescent images were photographed by an Olympus IX83 confocal microscope.
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