The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/100 - 1/250.
1/1000 - 1/10000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).
1/10 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Use at an assay dependent concentration. PubMed: 25058152
Is unsuitable for IHC-P or IP.
Adapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathway. 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.
Belongs to the 14-3-3 family.
Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Overlay histogram showing SH-SY5Y cells stained with ab92311 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab92311, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Western blot - 14-3-3 epsilon antibody [EPR3918] (ab92311)
All lanes : Anti-14-3-3 epsilon antibody [EPR3918] (ab92311) at 1/1000 dilution
Lane 1 : Human fetal brain cell lysate Lane 2 : SH-SY5Y cell lysate Lane 3 : 293T cell lysate Lane 4 : SW480 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : goat anti-rabbit HRP at 1/2000 dilution