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Sheep antibody to alpha synuclein (116-131): affinity purified


Catalogue No. S-075-50
Description Alpha synuclein is an abundant 140 amino acid neuronal protein, expressed primarily at presynaptic terminals in the central nervous system. Alpha synuclein has been associated with several neurodegenerative diseases. A point mutation in the gene coding for the alpha-synuclein protein was the first discovery linking this protein to a rare familial form of Parkinson's disease (PD). Subsequently, other mutations in the alpha-synuclein gene have been identified in familial PD. The aggregated proteinaceous inclusions called Lewy bodies found in PD and cortical Lewy body dementia (LBD) were discovered to be predominantly alpha-synuclein. Aberrant aggregation of alpha-synuclein has been detected in an increasing number of neurodegenerative diseases, collectively known as synucleopathies. Alpha-synuclein exists physiologically in both soluble and membrane-bound states, in unstructured and alpha-helical conformations, respectively. The physiological function of alpha-synuclein appears to require its translocation between these subcellular compartments and interconversion between the 2 conformations. Abnormal processing of alpha-synuclein is predicted to lead to pathological changes in its binding properties and function.
Batch No. See product label
Unit size 50 ug
Antigen A synthetic peptide (CMPVDPDNEAYEMPSEE) of human alpha synuclein protein (aa: 116-131) conjugated to diphtheria toxoid has been used as the immunogen.
Other Names Non-A beta component of AD amyloid; Non-A4 component of amyloid precursor; NACP
Accession SYUA_HUMAN
Produced in Sheep
Purity Affinity purified and dialysed against phosphate buffered saline (PBS).
Applications Immunohistochemistry (IHC): 1-4 ug/mL (paraffin sections)
Western Blotting (WB): 0.5 - 2.0 ug/mL. Fixing of proteins on membrane with 0.4% formaldehyde (30 min at room temperature) recommended, see Lee & Kamitani, 2011.
Flow Cytometry: 2 ug antibody per ~10^6 cells, methanol-fixed.
Immunocytochemistry (ICC): 1-4 ug/mL, 4% formaldehyde-fixed cells.

Biosensis recommends optimal dilutions/concentrations should be determined by the end user.
Specificity This antiserum specifically detects alpha synuclein.
Cross-reactivity This antibody is known to react with alpha synuclein of human, mouse, rat and other rodents.
Blast it If you would like to use this product in another species other than those specified here, or to see the shared identity between the immunogen used here in different speices and/or other molecules, simply copy the immunogen (from the Immunogen field) and paste it HERE and blast/format it. Note that, antisera raised against synthetic peptides are quite often very specific for that peptide ie, only one single amino acid difference may be enough to restrict the specificity to a particular molecule. Regardless, you can always contact us if you need assistance with this.
Form Lyophilised
Reconstitution Reconstitute in 50 uL of sterile water. Centrifuge to remove any insoluble material.
Storage Store lyophilized antibody at 2-8C. After reconstitution keep aliquots at -20C for a higher stability, and at 2-8C with an appropriate antibacterial agent. Glycerol (1:1) may be added for an additional stability. Avoid repetitive freeze/thaw cycles.
Expiry Date 12 months after purchase
Specific References 1. Kuo Y.M. et al. (2010) Extensive enteric nervous system abnormalities in mice transgenic for artificial chromosomes containing Parkinson disease-associated alpha-synuclein gene mutations precede central nervous system changes. Hum Mol Genet. May 1;19(9):1633-50
2. Pelkonen A. et al. (2010) Stimulated dopamine overflow and alpha-synuclein expression in the nucleus accumbens core distinguish rats bred for differential ethanol preference. J Neurochem. 2010 Aug;114(4):1168-76.
3. Alves et al. (2008) Striatal and nigral pathology in a lentiviral rat model of Machado-Joseph disease Hum Mol Genet. 2008 Jul 15;17(14):2071-83.
References 1. Lundvig et al, Brain Res Mol Brain Res 134, 3-17 (Mar 24, 2005).
2. Bennett, Pharmacol Ther 105, 311-31 (Mar, 2005).
3. Vekrellis, et al., Mol Neurobiol 30, 1-21 (Aug, 2004).
4. Martin, et al., Protein Pept Lett 11, 229-37 (Jun, 2004).
5. Doherty, et al., Acta Neuropathol (Berl) 107, 169-75 (Feb, 2004).
6. Goedert, Curr Opin Genet Dev 11, 343-51 (Jun, 2001).
7. Saito, et al., J Neurol Sci 177, 48-59 (Aug 1, 2000).
8. Lücking C.B. et al., Cell Mol Life Sci. 2000 Dec;57(13-14):1894-908.
9. Kahle, et al., Ann N Y Acad Sci 920, 33-41 (2000).
10. Clayton, et al., Trends Neurosci 21, 249-54 (Jun, 1998).
Images (click to zoom)
Sheep antibody to alpha synuclein (116-131): affinity purified Immunohistochemical staining (paraffin-embedded tissue) of glial inclusions in white matter from a case of multiple system atrophy using sheep polyclonal to human alpha synuclein, catalogue number S-075-50.
Sheep antibody to alpha synuclein (116-131): affinity purified Analysis of α-synuclein expression in formalin-fixed paraffin-embedded (FFPE) human Substantia Nigra tissue by Immunohistochemistry (IHC) using sheep antibody to α-synuclein (S-075-50, 4 ug/mL). Sections were cut to 4 um thickness and antigen retrieval performed using citrate buffer, pH 6, for 20 minutes. Primary antibody binding was visualized with a donkey anti-sheep-HRP conjugated secondary antibody (1:100) and 3’3-diaminobenzidine (DAB, brown). Cell nuclei were stained with a Hematoxylin nuclear counterstain (blue).
Sheep antibody to alpha synuclein (116-131): affinity purified Immunohistochemical staining of lewy body in Parkinsons diseased human brain tissue using sheep polyclonal to human alpha synuclein, catalogue number S-075-50.
Sheep antibody to alpha synuclein (116-131): affinity purified Analysis of alpha-synuclein expression by Flow Cytometry (A), Immunocytochemistry (B) and Western Blotting (C). The Flow Cytometry histogram (A) demonstrates presence of alpha-synuclein (blue line) in human SHSY-5Y neuroblastoma cells. Specific alpha-synuclein immunoreactivity (IR) is also observed in SHSY-5Y cells by Immunocytochemistry (B1, green), while under control conditions (B2), alpha-synuclein-IR is absent. In rodent tissue, the antibody detects α-synuclein monomer (~15 kDa) and a band at ~50 kDa on the blot, which most likely represents multimeric α-synuclein protein, which has been described by Shen et al., 2015, and others. Both bands are α-synuclein antibody specific as demonstrated by blocking with immunizing peptide. Recombinant human α-synuclein protein (100 ng) was run as control. Flow Cytometry method: Fixing and permeabilization: Absolute methanol (10 minutes on ice) and 0.1% Tween-20 in PBS; Blocking: 200 μg/mL sheep IgG; Primary antibody: sheep antibody to alpha-synuclein (S-075-50, 2 μg per ~10^6 cells, blue) for 30 minutes at room temperature; Secondary antibody: Goat anti-sheep, PE-labeled (1:100 dilution), 20 minutes in dark at room temperature. Negative control: Normal sheep IgG (S-1754-500, black). Data and results were generated using Orflo MoxiflowTM instrument and protocols. Immunocytochemistry method: Fixed (4% formaldehyde), permeabilized, and blocked (10% normal horse serum, 0.1% Triton X100) cells were incubated with alpha-synuclein antibody S-075-50 (2 μg/mL, green) for 1 hour. Primary antibody binding was visualized with a secondary donkey anti-sheep-CF488A antibody (4 μg/mL, 1 hour incubation). Cell nuclei were stained with Hoechst dye (blue). Control cells were stained following the same protocol, substituting the primary antibodies with normal sheep IgG (S-1754-500). Magnification: 100x. Western Blotting method: Protein loading amount: 50 μg per lane; Transfer: Towbin’s buffer with 20% methanol, semi-dry; Blocking: 5% skim milk; Primary antibody: 1 μg/mL, overnight incubation at 2-8°C; Secondary antibody: anti-sheep-HRP (1:6000); Detection: Chemiluminiscence. Proteins were fixed on membrane (0.4% formaldehyde, 30 minutes at room temperature) after transfer as described by Lee & Kamitani, 2011.
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