Laboratorio Universitario di Ricerca Medica, Univsersità degli Studi di Verona
No.
#100205
Date
27.11.2016
Antigène
Ga16
Numéro du lot
82100
Application validée
Western Blotting
Contrôle positif
QGP1 and PT45 lysates
Contrôle négative
MIA PaCa-2 cells lysate; PT45 shRNA knock-down cell lysate
Conclusion
Validation Images
Immunoblot in TBST using milk 5% as blocking agent; see protocol for details. A. Lysate from pancreatic cancer cell lines was loaded as indicated. B. Lysate from PT45 cells, untreated in lane 1 and treated with shRNA specific for GNA15 in lane 2.
Reveal protein bands using Luminata Forte Western HRP substrate (Millipore, WBLUF0500) as substrate and Syngene G-box to image light emission.
Strip membranes (2% SDS, 62.5mM TrisHCl pH6.8, 0.8% beta-mercaptoethanol) and subsequently incubate with a beta-actin loading control antibody (Santa Cruz clone (C4), sc-47778, lot D0108) diluted 1:1000 and secondary anti-mouse IgG HRP antibody (Sigma-Aldrich, A2554) diluted 1:10000.
Wash and reveal the protein as described above.
Notes
Passed. The inhibition proves that the antibody is specific for GNA15 and it does not recognize one of the more abundant and ubiquitous homologs, GNAQ or GNA11.
Format
Lyophilized
Stock
4 °C
Antigène
Ga16
Sujet
Ga 16 is the only hetero-trimeric G proteins with a restricted expression pattern in hematopoietic cells. Differentiation of promyelocyticcells leads to decreased expression of Ga 16. This G protein is known to couple a large number of G protein-coupled receptors to PLC-ß and can lead to production of the secondary messengers diacylglycerol and inositol phosphates. Targeted deletion of Ga 15, a mouse orthologue of human Ga 16, causes reduced C5a receptor signaling. Ga 16serves as a marker, in addition to CD34, for hematopoietic progenitor cells.