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Immunofluorescence analysis of Synuclein-alpha was done on 70% confluent log phase SH-SY5Y 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 Mouse Monoclonal Antibody (AHB0261) at 1ug/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 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® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant human alpha-synuclein.|
|Storage buffer||PBS, pH 7.2, with 0.1% BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1 µg/ml|
|Immunofluorescence (IF)||1 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:50|
|Western Blot (WB)||Assay Dependent|
* 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.
|ELISA (ELISA)||See 1 publications below|
A suggested positive control for this product is SHSY-5Y cells. It will not cross-react with mouse or rat.
Alpha-Synuclein belongs to the Synuclein family, which also includes beta and gamma Synuclein, and is predominantly expressed in neurons, concentrated at synaptic terminals. Synucleins may be involved in the regulation of dopamine release and transport and the modulation of synaptic vesicle function. Alpha-Synuclein is a small, presynaptic protein that is the major nonamyloid component of the pathological inclusions characteristic of a wide range of neurodegenerative disorders, collectively known as synucleinopathies. It is involved in the formation of SNARE complexes. Most significantly, aggregated Alpha-Synuclein is one of the major components found in the Lewy bodies that occur in Parkinson's disease (PD) and other neurodegenerative disorders. Mutation of the alpha-synuclein gene is associated with familial forms of PD. Alpha-Synuclein is known to reduce the fibrillization of the microtubule-associated protein, tau, to bind various targets such as 14-3-3, protein kinase C, synphilin-1, Elk and Tat-binding protein 1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Using gastrocnemius sEMG and plasma α-synuclein for the prediction of freezing of gait in Parkinson's disease patients.
AHB0261 was used in ELISA to investigate the use of surface electromyograph and plasma α-synuclein levels as predictors of the freezing of gait seen in patients with Parkinson's disease.
|Wang XY,Kang WY,Yang Q,Zhang LY,Chen SD,Liu J||PloS one (9:null)||2014|
alpha; Alpha-synuclein; NACP; non A-beta component of AD amyloid; Non-A beta component of AD amyloid; Non-A4 component of amyloid precursor; SYN; synuclein; synuclein alpha-140; synuclein, alpha (non A4 component of amyloid precursor); SYUA
NACP; PARK1; PARK4; PD1; SNCA