Immunofluorescent analysis of SNCA was performed on 70% confluent log phase HeLa 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 ABfinity™ SNCA recombinant rabbit oligoclonal antibody (Product # 710110) at a dilution of 1:1000 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 goat anti-rabbit IgG secondary antibody (Product # A11008) 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 (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e is a control without primary antibody. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 117–125 of human alpha synuclein|
|Contains||0.09% 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 *|
|Western Blot (WB)||1:500-1:5000|
* 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.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale and purified with Protein A.
ABfinity™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
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.