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 monoclonal antibody (Product # 701085) 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) and Panel d is a merged image showing nuclear localization and panel e is a control without primary antibody. The images were captured using a Nikon microscope at 20X magnification.
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Peptide corresponding to amino acids 117–125 of human synuclein, alpha|
|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 *|
|Flow Cytometry (Flow)||1:100-1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:1000|
|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.
|Western Blot (WB)||See 2 publications below|
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™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
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.
Nonaggregated ¿-synuclein influences SNARE-dependent vesicle docking via membrane binding.
701085 was used in western blot to examine the impact of nonaggregated α-Syn on SNARE-dependent liposome fusion.
|Lai Y,Kim S,Varkey J,Lou X,Song JK,Diao J,Langen R,Shin YK||Biochemistry (53:3889)||2014|
¿-Synuclein as an intrinsically disordered monomer--fact or artefact?
701085 was used in western blot to study whether alpha-synuclein exists as an intrinsically disordered monomer
|Coelho-Cerqueira E,Carmo-Gonçalves P,Pinheiro AS,Cortines J,Follmer C||The FEBS journal (280:4915)||2013|