14-3-3 proteins regulate many cellular processes relevant to cancer biology, notably apoptosis, mitogenic signaling and cell-cycle checkpoints. Seven isoforms comprise this family of signaling intermediates, denoted 14-3-3 b, g, e, z, h, q and s. 14-3-3 proteins form dimers that present two binding sites for ligand proteins, thereby bringing together two proteins that may not otherwise associate. These ligands largely share a 14-3-3 consensus binding motif and exhibit serine/threonine phosphorylation. 14-3-3 proteins function in broad regulation of these ligand proteins; by cytoplasmic sequestration, occupation of interaction domains and import/export sequences, prevention of degradation, activation/repression of enzymatic activity, and facilitation of protein modification. Loss of expression contributes to a vast array of pathogenic cellular activities.
Background References
1. Scheibner, KA. et al. 2012. MiR-27a functions as a tumor suppressor in acute leukemia by regulating 14-3-3θ. PLoS ONE. 7: e50895.
2. Song, Y. et al. 2012. Expression of 14-3-3γ in patients with breast cancer: correlation with clinicopathological features and prognosis. Cancer Epidemiol. 36: 533-536.
Sequence Similarity
Belongs to the 14-3-3 family.
Tissue Specificity
Highly expressed in brain, skeletal muscle, and heart.
Western blot analysis of 14-3-3 gamma on different lysates with Rabbit anti-14-3-3 gamma antibody (ET1612-9) at 1/2,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-14-3-3 gamma KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 28 kDa Observed band size: 28 kDa
Exposure time: 10 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 (ET1612-9) at 1/2,000 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.
Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-14-3-3 gamma antibody (ET1612-9) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-9) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue with Rabbit anti-14-3-3 gamma antibody (ET1612-9) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-9) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue with Rabbit anti-14-3-3 gamma antibody (ET1612-9) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-9) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of 14-3-3 gamma was done on K562 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-9, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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