Purification notesPurified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using i) a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated paxillin, and ii) a generic tyrosine phosphorylated peptide to remove antibody that is reactive with phosphotyrosine, irrespective of the sequence. The final product is generated by affinity chromatography using a paxillin-derived peptide that is phosphorylated at tyrosine 31.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.5 - 1 µg/ml. Predicted molecular weight: 68 kDa.
Use at an assay dependent concentration. PubMed: 24811250
FunctionCytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion).
Sequence similaritiesBelongs to the paxillin family. Contains 4 LIM zinc-binding domains.
Post-translational modificationsPhosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens.
Western blot - Paxillin (phospho Y118) antibody (ab4833)
Predicted band size : 68 kDa
Peptide Competition: Extracts prepared from NMµMG cells transfected with EGFP-tagged paxillin were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4oC, then were incubated with 0.75 µg/mL ab4833 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphotyrosine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab4833 blocks the antibody signal, thereby demonstrating the specificity of the antibody.
References for Anti-Paxillin (phospho Y118) antibody (ab4833)
This product has been referenced in:
Wang H et al. Inhibition of tumor vasculogenic mimicry and prolongation of host survival in highly aggressive gallbladder cancers by norcantharidin via blocking the ephrin type a receptor 2/focal adhesion kinase/paxillin signaling pathway. PLoS One9:e96982 (2014).
Read more (PubMed: 24811250) »
Lange K et al. Combined lysophosphatidic acid/platelet-derived growth factor signaling triggers glioma cell migration in a tenascin-C microenvironment. Cancer Res68:6942-52 (2008).
Read more (PubMed: 18757408) »