Rabbit anti cdc42

Whole and nuclear proteins were prepared as described previously.^{2 (link)} An aliquot of cell lysates containing 30 μg of protein was separated on 10% sodium dodecyl sulfate–polyacrylamide gels, and then transferred to polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). After blocked by 5% non-fat dry milk for 1 h at room temperature, membranes were incubated overnight at 4 °C with the following primary antibodies: rabbit anti-YAP (Cell Signaling Technology, Danvers, MA, USA, 1:1000), rabbit anti-Histone (Cell Signaling Technology, 1:3000), rabbit anti-Cdc42 (Santa Cruz, Dallas, TX, USA, 1:500), rabbit anti-Nwasp (Santa Cruz, 1:500), rabbit anti-Bax (Santa Cruz, 1:500), rabbit anti-Bcl-2 (Cell Signaling Technology, 1:1000), rabbit anti-GAPDH (Bioworld Technology, Nanjing, China, 1:10 000). The membranes were then incubated with anti-rabbit IgG (Jackson Immuno Research, West Grove, PA, USA, 1: 4000) at room temperature for 1h. Finally, membranes were treated with ECL reagents (Advansta, Menio Park, CA, USA), followed by exposing to X-ray film (Kodak, Rochester, NY, USA). The bands of the resulting autoradiographs were quantified densitometrically using Bandscan software. Protein expression was quantified as the ratio of specific band to Histone (nuclear fractions) or GAPDH.

Cdc42 and RhoA biosensors were kind gifts from Dr. Louis Hodgson (Albert Einstein College of Medicine). RhoA/Rac1/Cdc42 Activation Assay Combo Kit was purchased from Cell BioLabs (Sand Diego, CA, USA). QCM Gelatin Invadopodia Assay was obtained from Millipore (Massachusetts, USA). Primary antibodies against Cdc42, StarD13, actin, vinculin, Arp2, cortactin (Mouse and Rabbit) and TKS5 antibodies were obtained from Abcam (Cambridge, UK). Primary mouse anti-TKS4 was obtained from Merck Millipore. Primary mouse anti-StarD13 and rabbit anti-Cdc42 were obtained from Santa Cruz Biotech. HRP-conjugated secondary antibodies were obtained from Promega (Wisconsin, USA). Fluorescent secondary antibodies Alexa Fluor 488-green and Alexa Fluor 594-red as well as Rhodamine-phalloidin stain were purchased from Invitrogen (Massachusetts, USA). DAPI stain, and cell proliferation reagent were acquired from Roche Diagnostics (Roche Ltd, Mannheim, Germany). Hiperfect transfection reagent, luciferase and human Flexi Tubes siRNA for luciferase, StarD13, Cdc42 and RhoA were obtained from Qiagen (Hilden, Germany). Lipofectamine LTX was from Waltham (Massachusetts, USA) and crystal violet was from SCP Science (Quebec, Canada).