The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Synucleins are small, soluble proteins expressed primarily in neural tissue and in certain tumors. The family includes three known proteins: alpha, beta and gamma synucleins.
Synuclein-alpha is primarily localized at the presynaptic region of axons and it may be involved in the regulation of dopamine release and transport. However, it can form filamentous aggregates that are the major non amyloid component of intracellular inclusions in several neurodegenerative diseases (synucleinopathies). Synuclein-alpha may induce fibrillization of the microtubule-associated protein tau and it reduces neuronal responsiveness to various apoptotic stimuli, leading to a decreased caspase 3 activation. Synuclein-alpha is involved in Parkinson's disease, Lewy body dementia, Brain iron accumulation type 1 (NBIA1, also called Hallervorden-Spatz syndrome) and it is a major non-amyloid component of the deposits associated with Alzheimer's disease.
Synuclein-beta may be a regulator of the SNCA aggregation process. It protects neurons from staurosporine and 6 hydroxy dopamine (6OHDA)-stimulated caspase activation that also involves p53 and contributes to restore the SNCA anti-apoptotic function abolished by 6OHDA.
Gamma synuclein is found primarily in the peripheral nervous system and retina. High levels of expression have been identified in advanced breast carcinomas suggesting a correlation between overexpression of SNCG and breast tumor development.
ICC/IF image of ab6176 stained HeLa cells. The cells were 4% formaldehyde 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 (ab6176, 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.