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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 (NGF differentiated) cells stained with ab27766 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab27766, 1µg/1x106) for 30 min at 22°C. The secondary antibody used was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/2000 dilution for 30 min at 22°C.
Isotype control antibody (black line) was Mouse IgG1 [15-6E10A7] (ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5,000 events were collected using a 50mW Blue laser (488nm) and 530/30 bandpass filter.
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