Tight Junction Protein 1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Members of this family are involved in epithelial and endothelial intercellular junctions. They each contain at least one PSD95/Dlg/ZO-1 (PDZ) domain, a Src homology 3 (SH3) domain, and an enzymatically inactive guanylate kinase domain. PDZ domains are 90-amino acid protein-protein binding domains that recognize at least a 3-residue peptide motif in the COOH termini of their binding partners. PDZ domain-containing proteins, like ZO-1, typically act as scaffolding proteins that organize membrane receptors and cytosolic proteins into multimeric signaling complexes often at the sites of cell-cell contact. The effectiveness and stability of the epithelial barrier depends on a complex of proteins composing different intercellular junctions, which include tight junctions, adherens junctions, and desmosomes. ZO-1 is a peripheral membrane protein bound on the cytoplasmic surface of junctional contacts and is expressed in all tight junctions regardless of their properties. ZO-1 immunoprecipitates with its family member ZO-2. ZO-1 was shown to undergo tyrosine phosphorylation during tight junction formation and remodeling. Two different isoforms of ZO-1, alpha-minus and alpha-plus, have been described, which result from alternative splicing of an mRNA encoded by a single gene. The ZO-1 alpha-plus contains an 80 amino acids motif called alpha which is not present in ZO-1 alpha-minus. The alpha-containing isoform is found in most epithelial cell junctions. The short isoform (ZO-1 alpha-minus) is found both in endothelial cells and the highly specialized epithelial junctions of renal glomeruli and Sertoli cells of the seminiferous tubules. This difference in distribution provides molecular distinction among tight junctions.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-10877

• Tight Junction Protein 1 Antibodies
• Tight Junction Protein 1 ELISA Kits
• Tight Junction Protein 1 Lysate
• Tight Junction Protein 1 Proteins