Stokes Research Core

HPLC rules

1. The HPLC system can be booked for usage in either the morning or afternoon shifts.
2. Instrument usage rate: $12.50 /hr for trained users.
3. A training checkout is required before operation of the machine.
4. Please have a PCF staff member review your method before you begin. This can help avoid loss of your precious protein.
5. If you have signed up for time, you must use it and be courteous of the next user. Morning users must clean out the lines for the afternoon users. Evening users must do the same, regadless of the next day’s schedule. If no one is signed up for the next day or if it is Friday evening, the system must be stored in methanol for long term use and acetonitrile (0.1%TFA) for short term use. Please see the PCF staff for instructions.
6. The morning shift begins at 8:30 am. Failure to load sample by 10:00 will result in forfeiture of the time block. ALLOW 1 HOUR FOR SYSTEM PREPARATION. Afternoon users must begin by 1:00 pm and inject their samples by 2:00 pm.
7. All samples and buffers must be filtered. Failure to do so will damage the machine and result in a service call.
8. Fill out a user form after using the machine and put in the sample submission form box.
9. Sign the log book and report all problems immediately.
10. FAILURE TO USE THE INSTRUMENT DURING A RESERVED TIME PERIOD WILL RESULT IN A FULL SERVICE CHARGE
    

CD Rules

1. Get trained before using!
2. BEFORE TURNING ON THIS INSTRUMENT, purge with nitrogen for at least 30 minutes at 25 psi (read on gauge attached to right side of the instrument). If you fail to purge, or if nitrogen flow stops while the CD is on, you will fry the mirrors.
3. Make sure that half a tank of nitrogen is present in the tank before using!
4. Cells must be checked out and returned, CLEAN, to facility staff.
5. The facility staff is able to assist with data analysis – please inquire if you wish to use this service.
6. If this instrument is found to have been mis-used, there will be a minimum $100 restoration fee.
   

Centrifuge Rules

1. Sign-up for this instrument MUST be done 24 hours in advance – either on the web or at the instrument calendar.
2. We ask that you contact PCF staff if you plan to reserve more than 2 days on the centrifuge.
3. Only individuals trained by either Beckman or this facility may operate the centrifuge – NO EXCEPTIONS.
4. The facility would prefer to run samples for new users: Please contact Steven Seeholzer to arrange to have experiments run.
5. After completion of an experiment, please fill out the user form!
6. All users are required to clean the cells exactly as described in the cleaning instructions posted by the sink in the centrifuge room. Improper care of the cells can result in serious instrument damage!
7. Be sure to place Sapphire cells in their proper location. Confusion of Quartz and Sapphire cells will cause substantial delay in your experiments and other anguish.
8. We encourage new applicants to discuss their experimental needs with the facility staff.
9. Please cooperate with the facility staff in keeping this instrument in optimal condition. The staff are here to ensure that the instrument is fully operational at all times. Therefore problems must be brought to our attention immediately.

FPLC Rules

1. All new users MUST receive proper training before operating.
2. Please sign up at least 24 hours in advance of using the instrument.
3. Instrument usage rate: $12.50 /hr for trained users.
4. A training checkout is required before operation of the machine.
5. The AKTA system can be booked for usage in either the morning or afternoon shifts.
6. Trained users are welcome to use the instrument if it is open and unreserved.
7. Please have a PCF staff member review your method before you begin. This can help avoid loss of your precious protein.
8. If you have signed up for time, you must use it and be courteous of the next user. Morning users must clean out the lines for the afternoon users. Evening users must do the same, regadless of the next day’s schedule. If no one is signed up for the next day or if it is Friday evening, the system must be stored in 20% ethanol. Please see the PCF staff for instructions.
9. The morning shift begins at 8:30 am. Failure to load sample by 10:00 will result in forfeiture of the time block. ALLOW 1 HOUR FOR SYSTEM PREPARATION. Afternoon users must begin by 1:00 pm and inject their samples by 2:00 pm.
10. If you have reserved the AKTA for the whole day, we suggest preparing buffers the day before and equilibrating the column the evening before, so that purification can begin early in the morning.
11. All buffers and samples must be filtered before being applied to the instrument. All buffer containers must be sealed with parafilm.
12. Leaks are not acceptable.
13. Users are responsible for their own buffers. Please do not use any organic solvents, other than ethanol (20%), with this instrument.
14. Upon completion of your experiment:
    a. Please rinse the system with (millipore) water upon completion of your experiment.
    b. Remove tubes from the Fraction collector.
    c. Sign the logbook
    d. Fill out a user-form
    e. Clean your syringes
    f. Tidy up the area
    g. Leave it as you found it.
15. Chromatography columns are available for use. Please see Steven Seeholzer for questions regarding cost. Columns loaned out must be restored as per staff instructions. Columns are available for renting (ie SEC columns).
16. Chromatography media is available to users at cost.
17. Please fill out the user form immediately following use! If you fail to do this, we will charge for a full day’s use .
18. Sign the log book and report all problems immediately.
19. FAILURE TO USE THE INSTRUMENT DURING A RESERVED TIME PERIOD WILL RESULT IN A FULL SERVICE CHARGE.
20. This instrument must be stored in 20% ethanol (or water if someone is using it after you or the next day) after all experiments – no exceptions. If the instrument is found to be mis-treated, there will be a $150 restoration charge.
21. The protein core staff are available to assist in column and media selection.
22. The protein core facility can assist with purifications. Please ask for
    help, if you need it!

SELDI (CIPHERGEN) RULES

1. This instrument is not for confirmation of simple mass spectra – especially of peptides and small proteins. Please submit these samples for MALDI analysis. Large proteins can be analyzed on SELDI. Please consult the staff if you have questions
2. We strongly recommend desalting all samples. Zip-tips are available to users at cost ($5/tip).
3. Only trained users may operate this instrument. Contact Steven Seeholzer for training (seeholzer@email.chop.edu or 267.426.5679)
4. All users MUST fill out a sample analysis form BEFORE operating the instrument – NO EXCEPTIONS.
5. All users are responsible for providing their own coated sample chips. If you need coated chips, the facility can provide them at cost. Quantities of 2 or more chips must be pre-ordered.
   << reusable gold sample chips are provided free of charge >>
6. Signup for this instrument must be done 24 hours in advance. Trained users are welcome to use the instrument if it is open and unreserved.
7. Logbook signatures and form completion are required – violation will result in termination of privileges.
8. Any problems with the instrument must be reported to core facility staff – IMMEDIATELY.
9. All special projects that require prolonged instrument usage must be cleared by the core facility staff prior to signing up.

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