Laboratorio Universitario di Ricerca Medica, Univsersità degli Studi di Verona
No.
#100205
Datum
27.11.2016
Antigen
Ga16
Chargennummer
82100
Validierte Anwendung
Western Blotting
Positivkontrolle
QGP1 and PT45 lysates
Negativkontrolle
MIA PaCa-2 cells lysate; PT45 shRNA knock-down cell lysate
Bewertung
Validierungsbilder
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.
Anmerkungen
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
Lagerung
4 °C
Target
Ga16
Hintergrund
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.