Bov (Bovine), Hu (Human), Ms (Mouse), Rt (Rat), Chk (Chicken), Rb (Rabbit), Sh (Sheep), Can (Canine), Ze (Zebrafish), African clawed frog
Mouse, Sheep, Rabbit, Goat
Immunoprecipitation (IP), Western blot / Immunoblot (WB), Paraffin Sections (P), Immunocytochemistry/Immunofluorescence (ICC/IF), Enzyme Immunoassay (E), Frozen Sections (C)
Background of 14-3-3 protein theta antibody
At least seven isoforms comprise the highly conserved 14-3-3 family of homo-and heterodimeric proteins that are abundantly expressed in all eukaryotic cells. These multifunctional proteins bind and modulate the function of a wide array of cellular proteins. They exist mainly as dimers with a monomeric molecular mass of approximately 30 kDa and have an acidic isoelectric point of 4.5-5. Although more than seven isoforms of 14-3-3 have been described, some redundancies have appeared upon sequencing. The 14-3-3s are thought to be key regulators of signal transduction events mediated through their binding to serine-phosphorylated proteins. By interacting with Cdc25C, 14-3-3 regulates entry into the cell cycle and through interaction with bad, prevents apoptosis.3 14-3-3 plays a dual role in Raf-1 activation. It helps to keep Raf-1 in an inactive state by binding to phophoserine-259 and the cysteine-rich domain (DRD). Interactions of 14-3-3 at the phosphoserine-621 may be required for Raf-1 activation and may function as an essential cofactor for Raf-1 kinase activity.3 Other proteins that have been shown to bind 14-3-3s include members of the protein kinase C family, Cbl, IRS-1, middle-T antigen, KSR, IGF-1 receptor, etc.3 Detection of 14-3-3 proteins in cerebrospinal fluid has been shown to be quite useful in the differential diagnosis of Creutzfeldt-Jakob disease and other prion diseases. The theta or tau isoform was initially isolated from T cells.1 It
Howlett KF, Sakamoto K, Garnham A, Cameron-Smith D, Hargreaves M. Resistance exercise and insulin regulate AS160 and interaction with 14-3-3 in human skeletal muscle. Diabetes. 2007 Jun;56(6):1608-14. Epub 2007 Mar 16. PMID: 17369524
Cell lysates of Jurkat (20ug) were resolved by SDS-PAGE, transferred to NC membrane and probed with anti-human 14-3-3 tau (1:1000). Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and an ECL detection system.
Western Blot: 14-3-3 theta Antibody [NBP1-71900]- Whole cell lysate (1 mg for IP, 20% of IP loaded) from HeLa cells. Affinity purified rabbit anti-14-3-3-theta antibody used for IP at 6 mcg/mg lysate. 14-3-3-theta was also immunoprecipitated by rabbit anti-14-3-3-theta antibody NBP1-71901, which recognizes a downstream epitope.
Western Blot: 14-3-3 theta Antibody [NBP1-71901]-Whole cell lysate from HeLa (5, 15 and 50 mcg for WB; 1 mg for IP, 20% of IP loaded), 293T (T; 50 mcg) and mouse NIH3T3 (M; 50 mcg) cells. Antibodies: Affinity purified rabbit anti-14-3-3-theta antibody used for WB at 0.1 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 6 mcg/mg lysate. 14-3-3-theta was also immunoprecipitated by rabbit anti-14-3-3-theta antibody NBP1-71900, which recognizes an upstream epitope.