All lanes : Anti-Paxillin (phospho S178) antibody [21HCLC] (ab277786) at 2 µg/ml

Lane 1 : MDA-MB-231 (human breast adenocarcinoma cell line) whole cell extract
Lane 2 : PC3 (human prostate adenocarcinoma cell line) whole cell extract
Lane 3 : U-87 MG (human glioblastoma-astrocytoma epithelial cell line) whole cell extract
Lane 4 : Tissue extracts of Rat Brain
Lane 5 : Mouse Lung whole cell extract

Predicted band size: 65 kDa

For immunofluorescence analysis, MDA-MB-231 cells were fixed and permeabilized for detection of endogenous Paxillin pS178 using Anti- Paxillin pS178 Recombinant Rabbit Multiclonal Antibody (ab277786, 2 μ/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal^™ Secondary Antibody, Alexa Fluor^® 488 conjugate (1/2000). Panel a) shows representative cells that were stained for detection and localization of Paxillin pS178 protein (green), Panel b) is stained for nuclei (blue) using SlowFade^® Gold Antifade Mountant with DAPI. Panel c) represents cytoskeletal F-actin staining using Alexa Fluor^® 555 Rhodamine Phalloidin (1/300). Panel d) is a composite image of Panels a, b and c clearly demonstrating nuclear and cytoplasmic localization of Paxillin pS178. Panel e) shows loss of signal by competition with the Paxillin pS178 peptide, demonstrating antibody specificity. Panel f) demonstrates no competition with the non-phosphorylated peptide. Panel μ) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.