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Novel and adapted molecular strategies to create and trace toxic proteins in neurodegenerative disease
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Permanent Link: http://ufdc.ufl.edu/IR00001404/00001
 Material Information
Title: Novel and adapted molecular strategies to create and trace toxic proteins in neurodegenerative disease
Physical Description: Conference Proceedings
Creator: Workman, Aron S.
Slunt-Brown, Hilda
Borchelt, David R.
Conference: UF Society for Neuroscience Brain Awareness Week
Publication Date: 2008
 Notes
Acquisition: Collected for University of Florida's Institutional Repository by the UFIR Self-Submittal tool. Submitted by Aron Workman.
General Note: Support and PI by David R. Borchelt. Radiometric assay by Hilda Slunt-Brown. All other figures and text by Aron S. Workman.
 Record Information
Source Institution: University of Florida Institutional Repository
Holding Location: University of Florida
Rights Management: All rights reserved by the submitter.
System ID: IR00001404:00001

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Novel and adapted molecular strategies to create and trace toxic proteins in neurodegenerative disease Aron S. Workman, Hilda Slunt-Brown, and David R. Borchelt Department of Neuroscience and the McKnight Brain Institute University of Florida, Gainesville, FL 32612 I. PCR based mutagenesis II. Pulse chase coupled with nonionic detergent extraction III. Inducible neural cells IV Detecting superoxide radical antioxidants by riboflavin nitro blue tetrazolium assay 6 12 18 24 30 36 42 48 WT D 101G D 101N III. Inducible neural cells Analytical Biochemistry colored formazan superoxide antioxidant Light No dox 1 g/ mL dox HEK293 pTet On N2a pR Tet On 34 +contrast mouse neuroblastoma (line N2a) human neuroblastoma (line SH SY5Y) Retinoic acid differentiation aworkman@ufl.edu www.aronw.com S1 P2 35Scysteine pulse chase NP 40 insoluble protein aggregates soluble proteins and oligomers et al. Nature Success minipreps Fail minipreps MATERIALS AND METHODS Cell culture and transfections. SOD 1 aggregate extraction and immunoblotting g S1 P2 35Scysteine pulse chase analysis. Superoxide anion scavenging activity assay 1x TEN with 0.5% NP -40, 0.25% SDS, 0.5% deoxycholate and 1% PI et al Analytical Biochemistry