Cells were plated on poly-D-lysine (Sigma, P8920) coated coverslips (Fisher, 12–545-100) in 12 well plates. Cells untreated or treated with 100 nM Iso for 45 minutes at 37°C. Cells fixed in 3.7% formaldehyde (Fisher, F79) in modified BRB80 (80 mM PIPES pH 6.8, 1 mM MgCl2, 1 mM CaCl2) for 20 minutes. For cells to be stained, cells were blocked in 3% BSA (Sigma, A7030), 0.1% Triton X-100 (Sigma, T8787) in PBS. Next, cells were stained using antibodies (Supplementary Table 6 ) and with 1 µg/ml DAPI (20 mg/ml stock in water, Sigma, D9542) or 1 μM DRAQ5 (Invitrogen, 62251) in PBS. Then the coverslips were mounted on glass slides with ProLong Gold Antifade mounting media (Invitrogen, P10144). Slides were stored for a minimum of 24 hours to allow mounting media to dry before imaging.
P8920
Manufactured by Thermo Fisher Scientific
The P8920 is a high-performance laboratory centrifuge designed for a wide range of applications. It features a compact and durable construction, accommodating a variety of rotor types and sample volumes. The centrifuge provides precise speed and temperature control to ensure reliable and reproducible results.
Automatically generated - may contain errors
2 protocols using p8920
1
Imaging Cells After Isoproterenol Treatment
2
Imaging Cells After Isoproterenol Treatment
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Cells were plated on poly-D-lysine (Sigma, P8920) coated coverslips (Fisher, 12–545-100) in 12 well plates. Cells untreated or treated with 100 nM Iso for 45 minutes at 37°C. Cells fixed in 3.7% formaldehyde (Fisher, F79) in modified BRB80 (80 mM PIPES pH 6.8, 1 mM MgCl2, 1 mM CaCl2) for 20 minutes. For cells to be stained, cells were blocked in 3% BSA (Sigma, A7030), 0.1% Triton X-100 (Sigma, T8787) in PBS. Next, cells were stained using antibodies (Supplementary Table 6 ) and with 1 µg/ml DAPI (20 mg/ml stock in water, Sigma, D9542) or 1 μM DRAQ5 (Invitrogen, 62251) in PBS. Then the coverslips were mounted on glass slides with ProLong Gold Antifade mounting media (Invitrogen, P10144). Slides were stored for a minimum of 24 hours to allow mounting media to dry before imaging.
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