The 14-3-3 family of proteins plays a key regulatory role in signal transduction, checkpoint control, apoptotic and nutrient-sensing pathways (1,2). 14-3-3 proteins are highly conserved and ubiquitously expressed. There are at least seven isoforms, beta, gamma, epsilon, sigma, zeta, tau and eta that have been identified in mammals. The initially described alpha and delta isoforms are confirmed to be phosphorylated forms of beta and zeta respectively (3). Through their amino-terminal alpha helical region, 14-3-3 proteins forms homo- or heterodimers that interact with a wide variety of proteins: transcription factors, metabolic enzymes, cytoskeletal proteins, kinases, phosphatases and other signaling molecules (3,4). The interaction of 14-3-3 proteins with their targets is primarily through a phospho-Ser/Thr motif. However, binding to divergent phospho-Ser/Thr motifs, as well as phosphorylation independent interactions has been observed (4). 14-3-3 binding masks specific sequences of the target protein, and therefore, modulates target protein localization, phosphorylation state, stability and molecular interactions (1-4). 14-3-3 proteins may also induce target protein conformational changes which modify target protein function (4,5). Distinct temporal and spatial expression patterns of 14-3-3 isoforms have been observed in develop- ment and in acute response to extracellular signals and drugs, suggesting that 14-3-3 isoforms may perform differ- ent functions despite their sequence similarities (4). Several studies suggest that 14-3-3 isoforms are differentially regulated in cancer and neurological syndromes (2,3).
Applications:
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution:
Western Blot: 1:1000
Immunoprecipitation: 1:50
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.