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Lewy bodies purified from patients suffering dementia with Lewy bodies
Alpha-synuclein is expressed predominantly in the brain, where it is concentrated in presynaptic nerve terminals. The deposition of the abundant presynaptic brain protein alpha-synuclein as fibrillary aggregates in neurons or glial cells is a hallmark lesion in a subset of neurodegenerative disorders. These disorders include Parkinson's disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy, collectively referred to as synucleinopathies. Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the progressive accumulation in selected neurons of protein inclusions containing alpha-synuclein and ubiquitin.
Our Abpromise guarantee covers the use of ab27766 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent concentration.|
|WB||1/100 - 1/1000. Predicted molecular weight: 14 kDa.|
|IHC-Fr||1/100 - 1/1000.|
|IHC-P||1/100 - 1/1000. Do not perform antigen retrieval.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|ICC/IF||Use at an assay dependent concentration.|
|ICC||Use at an assay dependent concentration.|
SH-SY5Y neuroblastoma cells stained for alpha-synuclein (green) using ab27766 in immunofluorescence. SH-SY5Y cells were fixed with paraformaldehyde, permeabilized with 0.5% Tween-20 and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with ab27766 (diluted at 1/300) for 1 hour. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody (diluted at 1/200).
Human neuroblastoma cells stained for alpha-synuclein (green) using ab27766 in immunofluorescence. The neuroblastoma cells were fixed with paraformaldehyde and incubated with ab27766 (used at 5 μg/ml) for 12 hours at 4°C. A FITC conjugated Goat anti-Mouse IgG secondary antibody was then used.
Overlay histogram showing PC12 cells stained with ab27766 (red line). The cells were fixed with 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 alpha-synuclein antibody (ab27766, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC12 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.
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