This is a knockout-validated antibody summary, based on the publication "Multiple roles of filopodial dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Western blot analysis of lysates from wild-type (WT) and myeloid-restricted (conditional) Cdc42 knockout (Cdc42 cKO; Cdc42fl/fl/LysM-Cre) mouse peritoneal macrophages, after cell sorting for F4/80+ (mouse macrophage marker positive) cells. Each lane has 280,000 cells. Courtesy by the Authors.

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: Cdc42

Catalog number: 2466

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues surrounding Lys135 of human Cdc42. Reacts with human, mouse, rat and bovine. It is predicted to reacts with Xenopus. Suitable for western blot.

Western blot

Cells were lysed using Cell Lysis Buffer (#9803; Cell Signaling Technology) containing: 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1 % Triton, 2.5 mM sodium pyrophosphate, 1 mM β- glycerophosphate, 1 mM Na3VO4 (phosphatase inhibitor) and 1 µg/ml leupeptin (protease inhibitor), supplemented with 5 mM NaF (phosphatase inhibitor), 10 µg/ml aprotinin (inhibitor of trypsin and related proteases), 10 µg/ml Pefabloc SC (proteinase K inhibitor) and 1 mM DTT, a reducing agent.

TBS containing 5 % bovine serum albumin and 0.1 % Tween 20 for 1 h at room temperature.

1:1,000 dilution overnight at 4 °C.

Horseradish peroxidase-conjugated goat anti rabbit IgG polyclonal antibodies (111-035-003; Jackson ImmunoResearch Laboratories).

SuperSignal West Pico chemiluminescence substrate (Perbio, Bonn, Germany).