Immunofluorescence analysis of 14-3-3 theta was performed using 70% confluent log phase MCF-7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with 14-3-3 theta Rabbit Polyclonal Antibody (Product # 51-0700) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, C. elegans, Human, Mouse, Rat, Xenopus|
|Published species reactivity||Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A 20 amino acid synthetic peptide derived from the N-terminus of the human 14-3-3 beta/alpha protein.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.1-1.0 ug/ml|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Western Blot (WB)||2 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is broadly reactive with members of the 14-3-3 family of proteins (~29-32 kDa). With the exception of a single conserved amino acid change, this sequence is identical in the delta and zeta isoforms. Three and four conserved amino acid differences are found in the tau and gamma isoforms, respectively.
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. 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. Other proteins that have been shown to bind to 14-3-3s include members of the protein kinase C family, Cbl, IRS-1, polyoma middle-T antigen, nitrate reductase, S-raf and the IGF-1 receptor. 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
14-3-3 proteins regulate Tctp-Rheb interaction for organ growth in Drosophila.
51-0700 was used in immunocytochemistry and western blot to analyze organ growth in drosophila regulating the Tctp-Rheb interaction by 14-3-3 proteins
|Le TP,Vuong LT,Kim AR,Hsu YC,Choi KW||Nature communications (7:null)||2016|
|Not Applicable||Not Cited||
14-3-3zeta is indispensable for aggregate formation of polyglutamine-expanded huntingtin protein.
51-0700 was used in western blot to investigate the contribution of 14-3-3zeta to the aggregation of nonnative, aggregation-prone proteins
|Omi K,Hachiya NS,Tanaka M,Tokunaga K,Kaneko K||Neuroscience letters (431:45)||2008|
|Human||Not Cited||C-terminal recognition by 14-3-3 proteins for surface expression of membrane receptors.||Coblitz B,Shikano S,Wu M,Gabelli SB,Cockrell LM,Spieker M,Hanyu Y,Fu H,Amzel LM,Li M||The Journal of biological chemistry (280:36263)||2005|