216403b
Laboratory Course
WiSe 19/20: Protein Analysis and Microsequencing
Christoph Weise, Werner Schröder
Comments
Inhalt / contents
lectures: advanced methods in protein sequence analysis
Dr. C. Weise: chris.weise@biochemie.fu-berlin.de
Dr. W. Schröder: WernerSchroe@zedat.fu-berlin.de close
- Protein separation by 1D and 2D-PAGE
- Electro-blotting of proteins onto PVDF membrane
- Electro-elution and crush-and-soak extraction of proteins from gels
- Generation of peptides by enzymatic and chemical cleavage (in solution, in gel and on membrane)
- Derivatization and identification of cysteine and phosphoserine residues in proteins
- Performic acid oxidation
- Amino-acid analysis (total acid hydrolysis of proteins)
- Peptide separation by RP-HPLC
- Desalting of proteins and peptides by HPLC and SEC
- Manual N-terminal sequencing of proteins (PITC, ladder sequencing)
- Mass-spectrometric analysis of proteins and peptides
- Database search concerning protein identification and homology
- Sequence analysis (secondary structure prediction, Chou & Fasman, and hydropathy plot, Kyte & Doolittle)
- Automatic synthesis of DNA and RNA oligomers and their purification by HPLC and gel electrophoresis
lectures: advanced methods in protein sequence analysis
Dr. C. Weise: chris.weise@biochemie.fu-berlin.de
Dr. W. Schröder: WernerSchroe@zedat.fu-berlin.de close