Cells were grown on a confocal dish and treated with the compounds for 24 h. Next, the cells were fixed in 10% formalin for 15 min, washed thrice with phosphate-buffered saline (PBS), treated with 0.5% (v/v) Triton X-100/PBS for 15 min, washed thrice with PBS, and, subsequently, blocked with 10% FBS/PBS at 25 °C for 30 min. Cells were incubated with mouse anti-β-tubulin antibodies (Santa Cruz Biotech, Dallas, TX, USA) at 4 °C for 12 h, washed thrice with PBS, incubated for 30 min at room temperature with anti-mouse Alexa 488 secondary antibodies (Cell Signaling technology, Danvers, MA, USA) as a molecular probe, and washed thrice with PBS. Next, they were incubated with PI (10 µg/mL) and RNase (10 µg/mL) at 37 °C for 1 h. Intracellular microtubules and mitotic spindle formation were observed using FluoView FV10i confocal microscope (Olympus, Tokyo, Japan).
Anti mouse alexa 488 secondary antibodies
Manufactured by Cell Signaling Technology
Sourced in United States
Anti-mouse Alexa 488 secondary antibodies are fluorescently labeled antibodies used to detect and visualize primary antibodies raised against mouse antigens in immunoassays and other applications. These antibodies bind specifically to mouse primary antibodies, allowing for the detection and localization of target proteins or other molecules of interest.
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Lab products found in correlation
2 protocols using anti mouse alexa 488 secondary antibodies
1
Visualizing Microtubule Dynamics in Cells
2
PPAR-γ Expression in Ac2F Cells and Adipocytes
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The Ac2F cells and adipocytes (after differentiation) were seeded into the confocal dishes overnight. The cells were treated with parecoxib (15, 30, and 60 μM) and rosiglitazone (10 μM) for 48 h. Then, the cells were fixed with formalin for 10 min. The permeabilization of cells was achieved by incubating the cells with 0.3% Triton X-100 for 15 min. Next, the cells were blocked using 10% FBS and incubated with a PPAR-γ antibody at 4°C overnight. The cells were incubated with anti-mouse Alexa 488 secondary antibodies (Cell Signaling Technology) for 30 min, and then PI (10 μg/mL) and RNase (10 μg/mL) reagents were added to the cells and incubated for 40 min. Finally, FluoView FV10i confocal microscope (Olympus, Tokyo, Japan) was used to analyze the stained cells.
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