The Immunogen: The 14.3.3 family of proteins were originally discovered as prominent protein spots on 2 dimensional gels. They are a family of 28-33kDa proteins which are often major proteins of the cytoplasm (1). They function as binding partners for phosphoserine and phosphthreonine sites in other proteins, though they have other binding partners which are not dependent on phosphorylation. These binding interactions are important in the regulation of molecules such as the signaling kinases in the MAP kinase pathway c-Raf and b-Raf, the proapoptotic molecules Bad and Bax and the cell cycle regulator Cdc25. There are 7 mammalian 14.3.3 proteins and they are normally expressed as homo or in some cases heterodimers. 14.3.3η or 14.3.3 eta is widely expressed and concentrated in the nervous system. The HGNC name for 14.3.3 is YWHAH, and an alternate name is tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein 1, obviously due to a role for this protein in activation of tyrosine and tryptophan hydroxylases. One result of this is that the HGNC name is YWHAH. The 14.3.3η protein accumulates in the CSF of patients suffering form Creutzfeld Jacob Disease, and can be used for the diagnosis of this disease (2). Furthermore, this protein binds α-synuclein in the Lewy bodies of Parkinson's disease and has been linked to early-onset schizopherenia (3,4). The HGNC name for this protein is YWHAH.

Left: Blots of crude HeLa cell homogenate blotted with MCA-3G12 (left lane) and various molecular weight standards (right lane- numbers indicate apparent SDS-PAGE molecular weight in kDa). The MCA-3G12 monoclonal binds strongly and cleanly to a band at about 28kDa. Right shows rat mixed neuron/glial cultures stained with MCA-3G12 antibody (red). Neuronal perikarya are very rich in 14.3.3η which has a diffuse cytoplasmic staining pattern. Blue is a DNA stain.

Antibody characteristics: MCA-3G12 is a mouse IgG1 class antibody, and was raised against full length recombinant 14.3.3η expressed in and purified from E. coli. MCA-3G12 is known to react with 14.3.3η from human, mouse and rat. Since 14.3.3η is highly conserved, it is likely that the antibody is effective on other species also.

Suggestions for use: Try at dilutions of 1:500-1:1,000 for immunofluorescence. For western blots try at 1:1,000-1:5,000. A suitable control tissue is rat spinal cord or peripheral nerve homogenate.

OMIM link: press here

References:

1. Morrison, DK. The 14-3-3 proteins: integrators of diverse signaling cues that impact cell fate and cancer development. Trends Cell Biol. 19:16-23 (2009).

2. Hsich G, Kenney K, Gibbs CJ, Lee KH and Harrington MG. The 14-3-3 brain protein in cerebrospinal fluid as a marker for transmissible spongiform encephalopathies. N Engl J Med. 335:924-30 (1996).

3: Ubl A, Berg D, Holzmann C, Krüger R, Berger K, Arzberger T, Bornemann A, Riess O. 14-3-3 protein is a component of Lewy bodies in Parkinson's disease-mutation analysis and association studies of 14-3-3 eta. Brain Res Mol Brain Res. 108(1-2):33-9 (2002).

4: Middleton FA, Peng L, Lewis DA, Levitt P, Mirnics K. Altered expression of 14-3-3 genes in the prefrontal cortex of subjects with schizophrenia. Neuropsychopharmacology 30(5):974-83 (2005).

Limitations: This product is for research use only and is not approved for use in humans or in clinical diagnosis.

Availability and Price: Available for shipping now, $200 US per aliquot of 100 microliters of purified antibody at a concentration of 1 mg/ml, enough for hundreds of experiments. For order form press here.

Use of Images or Text: The contents of this page are available for modification and reuse under the terms of the Creative Commons Attribution/Share-Alike License 3.0 and the GNU Free Documentation License, unversioned with no invariant sections, front-cover texts, or back-cover texts. These licences permit modification and reuse, even commercially, as long as authorship credit and a link to this page is given.

©EnCor Biotechnology Inc. .