Protein

The Protein Microanalysis Facility provides "de-novo" N-terminal protein/peptide sequencing, internal sequencing / peptide mapping, amino acid composition analysis, peptide synthesis and mass spectrometry (ESI-MS, LC-MS and MALDI-TOF-MS). The ESI-MS allows for the soft ionization of peptides, proteins and phospholipids to determine accurate masses. Samples can be sprayed in directly or on-line when connected to an analytical HPLC. The combined use of chemical and enzymatic methods, such as in-gel-digestion of proteins from gel or PVDF pieces, together with bioinformatics software and public database searches allows for protein identification via mass or sequence tags using either MALDI or ESI. LC, HPLC and capillary electrophoresis are available for preparative and analytical runs. Two protein sequencers (Beckman LF 3000 ABI 477), an amino acid analyzer (ABI 420H) a peptide synthesizer (ABI 430), a capillary electrophoresis system (Beckman PACE 5000), an analytical (HPLC system (Beckman System Gold), an Electrospray-mass spectrometer (Bruker Esquire) and a MALDI-TOF mass spectrometer (Perseptive Biosystems Voyager) are operated in the Facility. The running of gels and electroblotting for sequencing also can be arranged.

Services and Fees
Amino Acid Analysis	$45 / run	Quantification of free amino acids and composition for peptides and proteins
Hydrolysis	$8 automated $10 manual	Peptides and proteins
Capillary electrophoresis	Starting at $45 for Each analysis	CE, CZE, ACE, IEF, SDS-CE analysis of Drugs, DNA, RNA, proteins & peptides
Mass Spectrometric Analysis (MS)	Starting at $45 for Each analysis	Mass analysis of biomolecules using matrix assisted laser desorption MS (MALDI)
HPLC; Proteins, Peptides & oligos	$45 each analytical run $80 preparative	Analysis and separations of biomolecules using liquid chromatography
Protein/Peptide Microsequencing	*$180 (1^st 10 cycles) additional $12 per cycle, $100 for 1^st 5 cycles, $100 set up.**	Edman based automated sequencing of protein and peptides, N-terminal and internal. *Only applicable if start of sequencer is scheduled before 11.00 am.
Protein Identification	$200 to 400 depending on state of sample and enzymes required	Generating of peptide ladders for proteomic research followed by isolation and sequencing of peptides. Includes peptide mapping, digests in gels, on membranes, or in solution.
Peptide Synthesis	$17.00 at the 20 mmole scale, desalted only. $20 to $50 per residue at the 0.25 mmole scale, $150 purification	Synthesis of short and long peptides, cyclic peptide, peptide nucleic acids (PNA’s), peptide mimics, and peptide conjugates
C-terminal Sequence Analysis	Call for info	Several approaches are available, e.g. Carboxypeptidase Y digest – MS, etc.
2D electrophoresis (to be added soon)	$100	Separation of complex protein mixtures
Native gel Electrophoresis	$25– 50	50 to 1000 kDa
-S-S- bridge analysis	$100 - $200	Folding in proteins and peptides
Phosphopeptide analysis	$60	Analysis of phosphorylated peptides
Cross-linking	*Call for info*	Cross-linking of biomolecules: protein-protein, protein-DNA
BIAcore	$15 / hour, $25 for setup	Real-time measurement of biomolecular interactions via surface plasmon resonance (SPR)
Flow Cytometer	$18 / hour	Qualitative analysis of a cell that passes through a focused laser beam
Luminometer	*No fee*	Chemiluminescence measurements in solution

Peptide Synthesis