Polyclonal Antibody to 14-3-3

Background
The 14-3-3 family of proteins are important regulatory molecules ubiquitiously expressed in all eukarryotic cells which bind to numerous signaling proteins in various pathways driving critical cellular pathways of apoptosis, differentiation and cell cycle (1-3). 14-3-3 theta specifically has also been shown to play an important regulatory role in the TLR2 signaling pathways (1) as a negative regulator of TLR2 ligand Pam3CySK4 induced NF-kB activation. 14-3-3 theta has previously been shown to interact with TLR4 ligand and MyD88 dependent phosphorylated PKC epsilon (3). 14-3-3 theta in the TLR4 signaling pathway is a positive regulator controlling release of IRF3 indcuded pro-inflammatory cytokines RANTES and IP-10 (3, 4). Currently identified by mass spec as part of the TLR2 signaling complex (1) and taken along with TLR4 data, a 14-3-3 theta antibody can be used to examine the different regulatory functions of 14-3-3 theta for different TLRs through its interaction with common or unique TLR signaling adaptor molecules in addition to MyD88 or PKC epsilon such as TRAM or TRIF allowing further clarification of TLR specific pathway regulation.

Antigen
A portion of amino acids 125-175 from human 14-3-3 was used as the immunogen for this antibody.

Application Notes
The amino acid sequence used as immunogen for this antibody is 100% homologous in human, mouse, panda, dog, chinese hamster, chicken, opossum, platypus, pig, frog, cat, orangutan, crab-eating macaque, and rhesus monkey, 94% in carolina anole, cow, zebrafish, and 89% homologous in frog, and rat.

Genebank Info (Protein)
NP_006817.1

Reference
1. Zhaoyuan Hou, Hongzhuang Peng, David E. White, Pu Wang, Paul M. Lieberman, Thanos Halazonetis, Frank J. Rauscher, III. 14-3-3 binding sites in the Snail protein are essential for Snail-mediated transcriptional repression and epithelial-mesenchymal differentiation. Cancer Res. 2010 June 1; 70(11): 4385–4393.
2. Catherine Johnson, Sandra Crowther, Margaret J. Stafford, David G. Campbell, Rachel Toth, Carol MacKintosh. Bioinformatic and experimental survey of 14-3-3-binding sites. Biochem J. 2010 April 1; 427(Pt 1): 69–78.
3. Diane L Bolton, Robert A Barnitz, Keiko Sakai, Michael J Lenardo. 14-3-3 theta binding to cell cycle regulatory factors is enhanced by HIV-1 Vpr. Biol Direct. 2008; 3: 17.
4. Kenny, EF and LAJ O’Neill 2008.  Signalling adaptors used by Toll-like receptors: An Update 43: 342-349.
5. Faisal A, et al 2008. The scaffold MYD88 acts to couple protein kinase epsion (PKC epsilon) to Toll-like Receptors J. Biol.Chem 283: 18591-18600.
6. Schuster TB et al 2011. Identification and functional characterization of 14-3-3 in TLR2 signaling. J Proteome Res. 10:4661-4670.
7. Obsilova, V et al 2008. 14-3-3 Proteins : a family of versatile regulators.   Physiol Rev. 57 (Suppl 3) s11-21.
8. Sridharan Rajagopalan, Robert S. Sade, Fiona M. Townsley, Alan R. Fersht. Mechanistic differences in the transcriptional activation of p53 by 14-3-3 isoforms. Nucleic Acids Res. 2010 January; 38(3): 893–906.
9. Yuki Matsui, Katsuya Satoh, Toshiaki Miyazaki, Susumu Shirabe, Ryuichiro Atarashi, Kazuo Mutsukura, Akira Satoh, Yasufumi Kataoka, Noriyuki Nishida. High sensitivity of an ELISA kit for detection of the gamma-isoform of 14-3-3 proteins: usefulness in laboratory diagnosis of human prion disease. BMC Neurol. 2011; 11: 120.
10. Yoshinari Asaoka, Fumihiko Kanai, Tohru Ichimura, Keisuke Tateishi, Yasuo Tanaka, Miki Ohta, Motoko Seto, Motohisa Tada, Hideaki Ijichi, Tsuneo Ikenoue, Takao Kawabe, Toshiaki Isobe, Michael B. Yaffe, Masao Omata. Identification of a Suppressive Mechanism for Hedgehog Signaling through a Novel Interaction of Gli with 14-3-3. J Biol Chem. 2010 February 5; 285(6): 4185–4194.