Rho gtpase antibody sampler kit

Whole proteins from the cultured cells were extracted according to the manufacturer’s protocol (RIPA) (Biyuntian, Shanghai, China), followed by concentration investigation. All samples were separated by SDS-PAGE and transferred following standard protocols. Antibodies to CD117 (Abcam, Cambridge, MA, USA) and FcεRI (Abcam) were used to identify mature mast cells. Rho-GTPase Antibody Sampler Kit (Cell Signaling Technology, Danvers, MA, USA), Phospho-MAPK Family Antibody Sampler Kit (Cell Signaling Technology) and Phospho-Stat Antibody Sampler Kit (Cell Signaling Technology), were used to detect the activation of signaling pathways in colon tumor cells. Antibody to cleaved caspase3 (Millipore, Temecula, CA, USA) was used to identify cell apoptosis. Antibody to β-tubulin (Cell Signaling Technology) was used to ensure the consistency of each cell lysate. Antibody to Pseudomonas Exotoxin A (Sigma-Aldrich Corp., St. Louis, MO, USA) and mouse anti-Human IgE antibody (Abcam) were used to identify the recombinant protein toxin. HRP-conjugated secondary antibodies were purchased from Cell Signaling and SuperSignal chemiluminescent reagent was purchased from Thermo Fisher Scientific (Waltham, MA, USA).
Protein expression was evaluated using Quantity One software according to immunoblotting band intensity.

Western blot was carried out as previously described [80 (link)]. Immunostaining was carried out using a goat monoclonal antibodies against Integrins: α1, α5, α6, αV and β3 (#ab34445, #ab150361, #ab20142, #ab179475, #ab75872, abcam) and Wnt/β-Catenin Activated Targets Antibody Sampler Kit (Cell Signaling # 8655S), β-Catenin Antibody Sampler Kit (Cell Signaling #2951S), Rho-GTPase Antibody Sampler Kit(Cell Signaling, #9968S), actin (1/1000, Cell signaling) and a secondary polyclonal mouse anti-goat antibody HRP conjugated (1/2000, cell signaling). Blots were developed using HRP and chemiluminescent peroxidase substrate (#CPS1120, Sigma). Data were collected using Geliance CCD camera (Perkin Elmer), and analyzed using ImageJ software (NIH).