Loredana Marchese
tel. 0382987260  loredana.marchese@unipv.it

The laboratory has extensive experience in amino acid analysis and N-terminal sequencing.

Amino acid analysis is carried out with precolumn derivatization technique using OPA and FMOC reagents. The laboratory is equipped for the acid hydrolysis of proteins, the preliminary treatment required for determination of amino acid composition of proteins or peptides. Moreover, determination of free amino acid present in biological fluids, pharmaceutical or food preparations is usually carried out. In general, 50 pmol/µl is the minimum concentration of sample required for the analysis and 50 µl is the minimum volume.

The automated Edman degradation is the technique used for sequencing peptides and proteins from the N-terminus. The analysis can be accomplished on proteins in solution. 600 pmoles is the minimum quantity of sample required to obtain a reliable sequence. However, purity of the sample, rather than the amount, is the determining factor for successful sequencing. Due to the limitations of the Edman degradation technique, peptides longer than around 20 residues can not be sequenced completely and a prior e-mail contact it’s due. Possible applications of these techniques are: analysis of purified or recombinant peptides/proteins, identification of unknown proteins, determinatio of amino acid composition of peptides/proteins, analysis of peptide-type drugs, analysis of food supplements based on amino acids-proteins-peptides.

Equipment

Jasco X-LC con rilevatore a fluorescenza collegato a HP Prodesk Core i5

Hewlett Packard modello Protein Sequencer G1000A, interfaced to a HP Kayak P III 500 MHz