All experiments were performed on a Zeiss Axioplan2 microscope (Carl Zeiss MicroImaging), with a 40x objective (Plan Neo, NA 0.75) and DeltaVision Image Restoration Microscope (Applied Precision). HCT116 cells were plated on glass coverslips (Fisher Scientific) in 6-well dishes 24 hours before fixation. Cells were exposed to DMSO only (vehicle control), ispinesib (50 and 100 nM) for 4 hours, or 50 ng/mL (166 nM) nocodazole for 14 hours at 37 °C in complete media. Cells were fixed for 10 min at 37°C in fix solution (4% formaldehyde, 0.2% Triton X-100, 10 mM EGTA, 1 mM MgCl2, 100 mM PIPES pH 6.8). Coverslips were washed 3 times with TBS-tx (TBS + 0.1% Triton X-100), blocked with AbDil (2% BSA in TBS-tx buffer) and incubated for 1 hr at room temperature with FITC-conjugated mouse anti-tubulin monoclonal antibody (Sigma # F2168; 1:2000 dilution in AbDil). Coverslips were washed three times in TBS-tx, and DNA was stained with Hoechst 33342 (Sigma; 1:10,000). Coverslips were mounted in 0.5% p-phenylenediamine (Sigma) in 20 mM Tris, at pH 8.8, with 90% glycerol and sealed with nail polish.
Plan neo
Manufactured by Cytiva
Plan Neo is a lab equipment product offered by Cytiva. It is a compact and versatile device designed for a range of laboratory applications. The core function of Plan Neo is to provide precise and efficient sample preparation and processing capabilities in a laboratory setting.
Automatically generated - may contain errors
Lab products found in correlation
P phenylenediamine, by Merck Group (2 mentions)
Glass coverslip, by Thermo Fisher Scientific (2 mentions)
Axioplan 2 microscope, by Zeiss (2 mentions)
Fitc conjugated mouse anti tubulin monoclonal antibody, by Merck Group (2 mentions)
Deltavision image restoration microscope, by Cytiva (2 mentions)
Hoechst 33342, by Merck Group (2 mentions)
2 protocols using plan neo
1
Microscopic Analysis of Microtubule Dynamics
2
Microscopic Analysis of Microtubule Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
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All experiments were performed on a Zeiss Axioplan2 microscope (Carl Zeiss MicroImaging), with a 40x objective (Plan Neo, NA 0.75) and DeltaVision Image Restoration Microscope (Applied Precision). HCT116 cells were plated on glass coverslips (Fisher Scientific) in 6-well dishes 24 hours before fixation. Cells were exposed to DMSO only (vehicle control), ispinesib (50 and 100 nM) for 4 hours, or 50 ng/mL (166 nM) nocodazole for 14 hours at 37 °C in complete media. Cells were fixed for 10 min at 37°C in fix solution (4% formaldehyde, 0.2% Triton X-100, 10 mM EGTA, 1 mM MgCl2, 100 mM PIPES pH 6.8). Coverslips were washed 3 times with TBS-tx (TBS + 0.1% Triton X-100), blocked with AbDil (2% BSA in TBS-tx buffer) and incubated for 1 hr at room temperature with FITC-conjugated mouse anti-tubulin monoclonal antibody (Sigma # F2168; 1:2000 dilution in AbDil). Coverslips were washed three times in TBS-tx, and DNA was stained with Hoechst 33342 (Sigma; 1:10,000). Coverslips were mounted in 0.5% p-phenylenediamine (Sigma) in 20 mM Tris, at pH 8.8, with 90% glycerol and sealed with nail polish.
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