It automates the following steps:
i) Washing of the gel slices with various buffers
ii) Reduction of disulfide bonds at 60 C
iii) Alkylation of cystine residues with iodoacetamide
iv) Removal of excess reagents
v) Equilibration with digest buffer
vi) Addition of digestion enzyme (optional up to 4 enzymes)
vii) Incubation at 37 C for digestion
viii) Elution of peptides from the gel matrix.
The major components of the system are:
i) Pipetting robot
ii) Dual syringe dilutor unit
iii) Work area with heatable reaction block, solvents and reagant rack, collection rack
iv) Built-in Touchpad and Liquid Crystal Display, single board computer, plus 3.5" disk drive and operating
software
v) Protective cover.
Enzymatic digestion is a standard technique used as part of the sample preparation process for a number of
analytical techiques which can include peptide mapping by liquid chromatography (LC) and capillary
electrophoresis (CE), but most commonly by MALDI-ToF or MS/MS mass spectrometry. The profile of the peptide
fragments generated by trypsin digestion (the most commonly used enzyme), can be considered to be a
fingerprint for each protein. The masses of each peptide fragment from a protein 'fingerprint' can be
accurately determined by mass spectrometry and these fragment sizes can be used to search against a variety
of databases for a 'match'. From this 'match' the identity of the protein may be determined.
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