The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesELISA: 1/40000.
ICC: Use at a concentration of 1 µg/ml.
IHC-P: 1/50 - 1/100.
WB: 1/500 - 1/1000. Detects a band of approximately ~28 kDa (predicted molecular weight: 28 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionAdapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathways. Binds to a large number of partners, usually by recognition of a phosphoserine or phosphothreonine motif. Binding generally results in the modulation of the activity of the binding partner.
Sequence similaritiesBelongs to the 14-3-3 family.
Post-translational modificationsThe delta, brain-specific form differs from the zeta form in being phosphorylated (By similarity). Phosphorylation on Ser-184 by MAPK8; promotes dissociation of BAX and translocation of BAX to mitochondria. Phosphorylation on Ser-58 by PKA; disrupts homodimerization and heterodimerization with YHAE and TP53. This phosphorylation appears to be activated by sphingosine. Phosphorylation on Thr-232; inhibits binding of RAF1.
Cellular localizationCytoplasm. Melanosome. Located to stage I to stage IV melanosomes.
This image shows paraffin-embedded human breast carcinoma tissue stained with ab51129 at a dilution of 1/100. Right hand image: tissue treated with immunising peptide; left hand image: untreated tissue
Western blot - 14-3-3 zeta antibody (ab51129)
All lanes : Anti-14-3-3 zeta antibody (ab51129)
Lane 1 : Extracts from 293 cells treated
with Forskolin (40nM, 30min) with no peptide Lane 2 : Extracts from 293 cells treated
with Forskolin (40nM, 30min) with immunising peptide
Predicted band size : 28 kDa Observed band size : 28 kDa
All lanes : Anti-14-3-3 zeta antibody (ab51129) at 1/500 dilution
Lane 1 : Recombinant Human 14-3-3 zeta protein (ab85268) at 0.1 µg Lane 2 : Recombinant Human 14-3-3 zeta protein (ab85268) at 0.01 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution developed using the ECL technique
ICC/IF image of ab51129 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab51129, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
References for Anti-14-3-3 zeta antibody (ab51129)
This product has been referenced in:
Simeone P et al. A unique four-hub protein cluster associates to glioblastoma progression. PLoS One9:e103030 (2014).
Read more (PubMed: 25050814) »
Ota S et al. Contribution of fetal ANXA5 gene promoter polymorphisms to the onset of pre-eclampsia. Placenta34:1202-10 (2013).
Read more (PubMed: 24140079) »