Generation and Characterization of Anti-α-Synuclein Antibodies

Mouse monoclonal antibodies 4A7 and 5D12 were generated against a synthetic peptide (CTKQGVAEAAGKTKEGVLYVGS) corresponding to amino residues 22–42 in human αS with an added N-terminal Cys generated as a service by Genescript and conjugated to Imject maleimide activated mariculture keyhole limpet hemocyanin (mcKLH; Thermo Scientific, Waltham, MA). Similarly, antibody 3H11 was generated with a synthetic peptide (CKTKEGVVHGVATVAEKTKEQ) corresponding to amino residues 43–62 in human αS and conjugated to mcKLC. These antibodies were generated by immunizing mice and procedures previously described in Dhillon et. al. 2017 (25 (link)). Other non-phospho-specific antibodies SNL4, 9C10, 33A-3F3 and 94–3A10 were previously described (25 (link),28 (link)). αS phospho-Ser129 (pSer129) antibodies 81A and LS4–2G12 were previously characterized (27 (link),29 (link),30 (link)) and EP1536Y was obtained from Abcam (Table 2).

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Dhillon J.K., Trejo-Lopez J.A., Riffe C., McFarland N.R., Hiser W.M., Giasson B.I, & Yachnis A.T. (2019). Dissecting α-synuclein inclusion pathology diversity in multiple system atrophy: implications for the prion-like transmission hypothesis. Laboratory investigation; a journal of technical methods and pathology, 99(7), 982-992.

Publication 2019

mcKLH EP1536Y

Antibodies Antibody Human Keyhole limpet hemocyanin Maleimide Mice Monoclonal antibodies Peptide Phospho specific antibodies

Corresponding Organization : University of Florida

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Variable analysis

independent variables

Mouse monoclonal antibodies 4A7 and 5D12 generated against a synthetic peptide (CTKQGVAEAAGKTKEGVLYVGS) corresponding to amino residues 22–42 in human αS with an added N-terminal Cys
Antibody 3H11 generated with a synthetic peptide (CKTKEGVVHGVATVAEKTKEQ) corresponding to amino residues 43–62 in human αS

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

positive controls

Other non-phospho-specific antibodies SNL4, 9C10, 33A-3F3 and 94–3A10 previously described
αS phospho-Ser129 (pSer129) antibodies 81A and LS4–2G12 previously characterized
αS phospho-Ser129 (pSer129) antibody EP1536Y obtained from Abcam

negative controls

Not explicitly mentioned

Annotations

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