Cdc42 Polyclonal Antibody

Protocol tips

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	-Rabbit (1:1000)

Protocol tips
-Rabbit (1:1000)

Publication protocol

For validation of CDC42 antibody specificity, Western blotting was performed on whole cell lysates of MCF-7, SKBr3 and MDA-MB231 human breast cancer cell lines (obtained from the American Type Culture Collection; Rockville, MD, USA) using CDC42 antibody (clone PA1-092) at 1:1000 dilution and fluorescent secondary antibodies at 1:15,000 were used (IR Dye 800CW donkey anti-rabbit and 680RD donkey anti-mouse, LI-COR Biosciences, UK). 5% milk (Marvel original dried skimmed milk, Premier Food Groups Ltd, St Albans, UK) was used for blocking. Mouse β-Actin (A5441, Sigma-Aldrich; Clone AC-15; Sigma, UK) at 1:5000 was used as a house-keeping protein. A protein ladder (PageRuler Plus Prestained Protein Ladder, ThermoScientific, Waltham, MA, USA) was included. The fluorescence was then detected using the LI-COR Odyssey Fc machine to visualise the bands, with wavelengths 600, 700 and 800.

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