At 3 days after differentiation, PLB-985 cells were spun down and resuspended in extracellular buffer (5 mM KCl, 125 mM NaCl, 1.5 mM CaCl2, 1.5 mM MgCl2, 10 mM Glucose, 20 mM HEPES, pH 7.4). 0.9 uM Control (Dharmacon, D-001206-14-05), Cdc42 (Dharmacon, M-005057-01-0005), and RhoA (Dharmacon, E-003860-00-0005) siRNA pools were mixed with the cell suspension and were introduced into cell by parallel electroporation using a custom-built 96-well electroporation device69 (link). Cells were then incubated 3 days more with differentiation media in 96 well plate. For western blot, whole-cell extracts were prepared and resolved by 4%-10% gradient SDS-PAGE. The proteins were then transferred to a nitrocellulose membrane. Primary antibodies, anti-Cdc42 (Cell Signaling Technology, 2462, 1:1,000), anti-RhoA (Cell Signaling Technology, 2117, 1:1,000), and anti-GAPDH (Cell Signaling Technology, 5174, 1:1,000), were incubated overnight at 4°C. Bound antibodies were visualized with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) and the ECL system (Millipore).
Cdc42
Manufactured by Horizon Discovery
Cdc42 is a Rho family GTPase that plays a key role in regulating various cellular processes, such as cell morphology, migration, and signaling. It functions as a molecular switch, cycling between an active GTP-bound state and an inactive GDP-bound state. Cdc42 is involved in a wide range of cellular activities, including cytoskeletal organization, vesicle trafficking, and gene expression.
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Lab products found in correlation
2 protocols using cdc42
1
Efficient Knockdown of Cdc42 and RhoA in PLB-985 Cells
2
Efficient Knockdown of Cdc42 and RhoA in PLB-985 Cells
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At 3 days after differentiation, PLB-985 cells were spun down and resuspended in extracellular buffer (5 mM KCl, 125 mM NaCl, 1.5 mM CaCl2, 1.5 mM MgCl2, 10 mM Glucose, 20 mM HEPES, pH 7.4). 0.9 uM Control (Dharmacon, D-001206-14-05), Cdc42 (Dharmacon, M-005057-01-0005), and RhoA (Dharmacon, E-003860-00-0005) siRNA pools were mixed with the cell suspension and were introduced into cell by parallel electroporation using a custom-built 96-well electroporation device69 (link). Cells were then incubated 3 days more with differentiation media in 96 well plate. For western blot, whole-cell extracts were prepared and resolved by 4%-10% gradient SDS-PAGE. The proteins were then transferred to a nitrocellulose membrane. Primary antibodies, anti-Cdc42 (Cell Signaling Technology, 2462, 1:1,000), anti-RhoA (Cell Signaling Technology, 2117, 1:1,000), and anti-GAPDH (Cell Signaling Technology, 5174, 1:1,000), were incubated overnight at 4°C. Bound antibodies were visualized with horseradish peroxidase-conjugated secondary antibodies (GE Healthcare) and the ECL system (Millipore).
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