216403b Laboratory Course

WiSe 21/22: Protein Analysis and Microsequencing

Jens Peter Fürste, Werner Schröder


Inhalt / contents

  • 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
theoretical: mass spectrometry in protein and nucleic acid analysis
lectures: advanced methods in protein sequence analysis

Dr. J. P. Fürste: fuerste@zedat.fu-berlin.de
Dr. W. Schröder: WernerSchroe@zedat.fu-berlin.de close

Subjects A - Z