Anti-His tag antibody, clone IJT.3B9

In this article, the magnetic nanospheres bonded to a polymer chelator have synthesized through a new strategy consisted of four parts: (1) synthesis and surface modification of nanoparticles with vinyl groups; (2) grafting of a water-soluble polymer having hydroxyl groups, by polymerization of the resulting monomer from the reaction of glycidyl methacrylate (GMA) and diethanolamine (DEA) on the surface of nanomagnetic particles; (3) by cerium(IV), conversion of hydroxyl groups of the synthesized polymer on the nanoparticles to radicals for subsequent grafting of the resulting monomer from reaction of GMA and iminodiacetic acid (IDA); and (4) in an ion-exchange process, nickel ions along the last synthesized polymer were immobilized. The prepared nanospheres were successfully used directly for purification of His-tagged xylanase from cell lysates. The separated proteins were subjected to enzyme assays and are observed well in the enzymatic activity.

OBJECTIVE: To explore the role of integrin alpha v in Angiotensin II (Ang II)-induced exocytosis and endocytosis of galectin-3 (gal-3) in vascular smooth muscle cells (VSMCs). MATERIALS AND METHODS: A primary culture of mouse VSMCs was established by the enzymatic digestion of aorta. Adeno-Cre was used to specifically knockdown integrin alpha v. VSMCs were treated with Ang II, LY294002 (inhibitor of AKT signaling pathway), and Bay11-7082 (inhibitor of nuclear factor-kappa B, NF-kappa B), respectively. Endocytosis of His-tagged gal-3 was analyzed by immunofluorescence. The Western blot was performed to detect the protein level in cell supernatant and lysate. RESULTS: Ang II increased the exocytosis of gal-3 and activated AKT and NF-kappa B signaling pathways. The knockdown integrin alpha v effectively decreased the activation of AKT and NF-kappa B signals and the exocytosis of gal-3 induced by Ang II, but it had a little effect on the endocytosis of gal-3. Ang II increased the phosphorylation of AKT and NF-kappa B through integrin alpha v. AKT is the upstream signal of the NF-kappa B signaling pathway. LY294002 or Bay11-7082 could decrease Ang II-induced exocytosis of gal-3 in VSMCs. CONCLUSIONS: Ang II, depending on integrin alpha v/AKT/NF-kappa B signaling pathway, induced the exocytosis of gal-3.