The 14-3-3 proteins are a family of proteins involved in the regulation of apoptosis, mitogenic signaling and cell-cycle checkpoints. The 14-3-3 proteins are thought to be key regulators of signal transduction events mediated through their binding to serine-phosphorylated proteins. Through binding Bad, 14-3-3 prevents apoptosis by sequestering Bad to the cytosol. The 14-3-3 proteins are also Adapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathway. They bind to a large number of partners, usually by recognition of a phosphoserine or phosphothreonine motif. Binding generally results in the modulation of the activity of the binding partner.
Background References
1. Carninci P., Kasukawa T., et al. Science 309:1559-1563(2005).
3. Fu, H., et al. 14-3-3 proteins: structure, function, and regulation. Ann. Rev. Pharmacol. Toxicol. 40: 617–47 (2000).
Sequence Similarity
Belongs to the 14-3-3 family.
Post-translational Modification
Isoform alpha differs from isoform beta in being phosphorylated (By similarity). Phosphorylated on Ser-60 by protein kinase C delta type catalytic subunit in a sphingosine-dependent fashion.; Isoform Short contains a N-acetylmethionine at position 1.
Western blot analysis of 14-3-3 alpha+beta on different lysates with Rabbit anti-14-3-3 alpha+beta antibody (0407-5) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-14-3-3 alpha+beta KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 28 kDa Observed band size: 28 kDa
Exposure time: 20 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (0407-5) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ICC staining 14-3-3 b/a in N2A cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS.
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