RAC1 Antibody

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Rabbit (1:500) -Soak PVDF membranewith pore size 0.45 um in methanol for 30 sec -Apply Wet transfersystems at 180mA for 90 min with Wet transfer buffer.

Protocol tips
-Rabbit (1:500) -Soak PVDF membranewith pore size 0.45 um in methanol for 30 sec -Apply Wet transfersystems at 180mA for 90 min with Wet transfer buffer.

Publication protocol

Protein samples were fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes (Millipore). After blocking with 5% non-fat milk for 2 h at room temperature, the membranes were incubated with primary antibodies against NKD1 (1:1000, #2262, CST), Rac1 (1:500, 66122-1-Ig, Proteintech), EZH2 (1:1000, #5246, CST), GAPDH (1:5000, AT0010,CMCTAG), β-Actin (1:1000, AT0001, CMCTAG), E-cadherin (1:1000, #14472, CST), Flag (1:1000, F1804, Sigma), and Myc (1:500, 60003-2-Ig, Proteintech) at 4 °C overnight. Subsequently, the blots were incubated with the corresponding secondary antibody labeled with horseradish peroxidase (Yeasen) at room temperature for 2 h. Protein bands were quantified by densitometry using Image-J software and compared to the reference protein. Experiments were repeated in triplicate independently.

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Manufacturer protocol

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