Immunofluorescence analysis of Synuclein-alpha + beta was done on 70% confluent log phase SHSY5Y cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Synuclein-alpha + beta Mouse Monoclonal Antibody (328200) at 1.5 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Mouse|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Recombinant human alpha- and beta-synuclein.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||1-2 ug/ml|
|Immunofluorescence (IF)||1-2 ug/ml|
|Immunohistochemistry (IHC)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 ug/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.
|Western Blot (WB)||See 1 publications below|
Synuclein family of proteins is abundantly expressed in neuronal cytosol and presynaptic terminals. In vertebrates they are encoded by three different genes. Synucleins have been specifically implemented in three major diseases: Alzheimer's (AD), Parkinson's PD) and breast cancer. In AD, a peptide derived from alpha synuclein forms an intrinsic component of plaque amyloid. In PD, an alpha synuclein accumulates in Lewy bodies. An allele of alpha synuclein has been linked to many familial cases of PD. In breast cancer increased expression of gamma synuclein correlates with the disease progression. Synucleins appear to be involved in the membrane plasticity in developing song control system of songbirds.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Mouse||Not Cited||Synaptic vesicle depletion correlates with attenuated synaptic responses to prolonged repetitive stimulation in mice lacking alpha-synuclein.||Cabin DE,Shimazu K,Murphy D,Cole NB,Gottschalk W,McIlwain KL,Orrison B,Chen A,Ellis CE,Paylor R,Lu B,Nussbaum RL||The Journal of neuroscience : the official journal of the Society for Neuroscience (22:8797)||2002|