| Products/Services Used | Details | Operation |
|---|---|---|
| Peptide Synthesis | A library of 70 overlapping peptides covering the entire sequence of the rat Panx2 (Uniprot accession number P60571) minus the four transmembrane domains was obtained from Genscript (Piscataway, NJ, USA). Peptides were 15 amino acids in length with a 7 amino acids overlap. A total of 100 µg of peptide was pre-incubated overnight at 4°C in 100 µL of dot-blot buffer (8 M Urea, 100 mM NaH2PO4, 10 mM Tris, pH 8.0) containing 10 µg bovine serum albumin. Peptides were then dot-blotted on nitrocellulose membrane (Bio-Rad, Hercules, CA, USA), washed with sodium phosphate buffered saline (PBS, pH 7.4), dried at 37°C and blocked for 1 h in milk solution (4% nonfat milk, 20 mM Tris, 150 mM NaCl pH 7.4). The membrane was then immunoprobed for 2 h at room temperature with the primary antibody followed by HRPO-conjugated secondary antibodies (Sigma, St. Louis, MO, USA) for 1 h at room temperature. | Get A Quote |
Pannexins (Panx) are proteins homologous to the invertebrate gap junction proteins called innexins (Inx) and are traditionally described as transmembrane channels connecting the intracellular and extracellular compartments. Three distinct Panx paralogs (Panx1, Panx2 and Panx3) have been identified in vertebrates but previous reports on Panx expression and functionality focused primarily on Panx1 and Panx3 proteins. Several gene expression studies reported that Panx2 transcript is largely restricted to the central nervous system (CNS) hence suggesting that Panx2 might serve an important role in the CNS. However, the lack of suitable antibodies prevented the creation of a comprehensive map of Panx2 protein expres... More
