Anti flag tag mouse monoclonal igg antibody

Lung tissues or treated cells were lysed in protein extraction buffer [50 mM Tris-Cl (pH 7.5), 150 mM NaCl, 1 mM EDTA (pH 8.0), 1% Triton X-100, and protease inhibitor cocktail (Roche Molecular Biochemicals, Indianapolis, IN, USA)] for 30 min on ice. Western blot analysis and co-immunoprecipitation assay were performed as previously described (22 (link)) using the following antibodies: anti-Smac rabbit monoclonal antibody (1:1,000; ab32023), anti-XIAP rabbit polyclonal antibody (1:2,000; ab21278), anti-β-actin rabbit polyclonal antibody (1:5,000; ab75186) (both from Abcam, Cambridge, UK); anti-GAPDH rabbit polyclonal IgG antibody (1:1,000, sc-25778; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA); anti-Flag tag mouse monoclonal IgG antibody (1:1,000, F1804; Sigma-Aldrich, St. Louis, MO, USA); anti-myc tag mouse monoclonal antibody (1:1000; 2276S; Cell Signaling Technology, Inc., Danvers, MA, USA); anti-ubiquitin rabbit monoclonal antibody (1:2000; ab140601; Abcam).

Lung cancer cells were fixed with 3% paraformaldehyde and permeabilized using 0.2% Triton X-100. Then the fixed cells were incubated with anti-Flag tag mouse monoclonal IgG antibody (1:1,000, F1804; Sigma-Aldrich). The secondary antibody is a labeled goat anti-mouse IgG antibody (A11001; Invitrogen, Carlsbad, CA, USA). Fluorescence confocal images were captured using a LSM 5 Pascal Laser Scanning Microscope (Carl Zeiss AG, Oberkochen, Germany) and Laser Scanning Microscope LSM PASCAL software (version 4.2 SP1).