* Allow all reagents to warm to room temperature for at least 30 minutes before opening.
* Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.
* Unused wells must be kept desiccated at 4 degree C in the sealed bag provided.
* Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.
* It is recommended that all samples and standards be assayed in duplicate or triplicate.
Background: Paxillin is a signal transduction adaptor protein and should not be confused with the neurotoxin paxilline.Glenney and Zokas (1989) used an antiphosphotyrosine antibody to identify proteins that are phosphorylated in Rous sarcoma virus-transformed chick embryo fibroblasts, and found a 76-kD protein that localizes to focal adhesions at the ends of actin-containing stress fibers in nontransformed cells. Turner et al. (1990) purified this protein from chicken gizzard smooth muscle, and named it paxillin ('paxillus' means 'small stake' or 'peg' in Latin) as a protein tethered to the membrane at focal adhesions.Paxillin migrates as a diffuse 65- to 70-kD band on SDS-PAGE. Salgia et al. (1995) stated that transmembrane integrin molecules connect the actin cytoskeleton to the extracellular matrix within focal adhesions.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion). Alternatively spliced transcript variants encoding different isoforms have been described for this gene. These isoforms exhibit different expression pattern, and have different biochemical, as well as physiological properties (PMID:9054445). [provided by RefSeq, Aug 2011]
Uniprot Description
PXN: a multi-domain cytoskeletal protein involved in actin-membrane attachment that localizes primarily to focal adhesion sites to the extracellular matrix. Phosphorylated by focal adhesion kinase (FAK) and is a component of integrin signaling. Its phosphorylation provides docking sites for recruitment of signaling molecules to focal adhesions. Binds in vitro to vinculin as well as to the SH3 domain of c-SRC and, when tyrosine phosphorylated, to the Crk SH2 domain. Interacts with GIT1, NUDT16L1/SDOS, PARVA and TGFB1I1. Component of cytoplasmic complexes, which also contain GIT1, ARHGEF6 and PAK1. Binds DDEF2. Interacts with unphosphorylated ITGA4. Interacts with RNF5.Three alternatively-spliced isoforms have been described. Isoform beta binds to focal adhesion kinase but weakly to vinculin. Isoform gamma binds to vinculin but only weakly to focal adhesion kinase.
Protein type: Adaptor/scaffold; Motility/polarity/chemotaxis; Cell adhesion; Cytoskeletal
Chromosomal Location of Human Ortholog: 12q24.31
Cellular Component: nucleoplasm; microtubule associated complex; focal adhesion; lamellipodium; cytoplasm; plasma membrane; stress fiber; cell cortex; cytosol
Molecular Function: integrin binding; protein binding; zinc ion binding; beta-catenin binding; BH4 domain binding; protein kinase binding; vinculin binding
Biological Process: epidermal growth factor receptor signaling pathway; lamellipodium biogenesis; focal adhesion formation; integrin-mediated signaling pathway; peptidyl-tyrosine phosphorylation; activation of MAPK activity; signal complex assembly; cytoskeleton organization and biogenesis; signal transduction; regulation of cell shape; branching morphogenesis of a tube; muscle contraction; transforming growth factor beta receptor signaling pathway; vascular endothelial growth factor receptor signaling pathway; cell adhesion