Resources

 

Perkin Elmer-Sciex prOTOF MALDI: This instrument is a reflectron time-of-flight (TOF) mass spectrometer which detects proteins and peptides spotted on disposable conductive plates in 96 and 384-well formats. The ionization method is MALDI (matrix assisted laser desorption and ionization) where a desalted protein sample is mixed with an organic acid matrix. The matrix forms crystals around the protein species when dried and transfers a positive charge by protonation after being excited by laser light. The prOTOF measures the intact masses of proteins and peptides from 500 Da to 200 kDa with resolutions up to 20,000 and sub-femtomolar sensitivities. The sensitivity of MALDI (and other mass spectrometers) generally increases as the mass of proteins or peptides decreases. The optimal mass range for this instrument is from 1-20 kDa. The novel design of the detection system allows for a single calibration for an entire plate or even a series of plates, which ensures extremely accurate mass measurements (<20 ppm for peptides less than 5000 Da) for a large number of samples.

 

Hamilton STARlet Liquid-Handling Robot: This is a pipetting workstation that will process all our protein samples in 96 and 384-well formats. It performs filtrations on a vacuum station, mixing on 3 independent shakers, solid-phase extraction by magnetic-bead separations, cooling of samples on a Peltier plate, and moves plates between stations with a set of gripper arms. The accuracy of pipetting (%CV’s < 2%) greatly improves the reproducibility of our assays.

 

Shimadzu Prominence HPLC: The HPLC allows the fractionation of very complex mixtures into much simpler portions. The Shimadzu system accommodates a wide range of flow rates - from 1 mL/min to 10 mL/min - so both preparative columns (including plasma depletion columns) with large protein loads and analytical columns with as little as 1 mg of protein can be run. The autoloader feature allows hands-free automatic injection of two chilled 96-well plates. The fraction collector is also cooled and accepts 96 and 384 well plates. The dual-wavelength UV-VIS detector is tunable and can be set between 190 and 700 nm.