Shared Facilities

Bio-Plex

Policies & Procedures

To use the core, please do the following:

  • Complete the “Authorization form.” Follow submission guidelines on the form.
  • Refer to “Procedures” below for information on sample collection. Additional instructions are included in each kit as well as on the Bio-Rad Web site (keyword: “Bio-Plex”) or the Millipore website (keyword “Milliplex”).
  • Refer to the Assay Builder Web site for information on designing custom assays, price quotes, Express Assays, x-Plex Assays, and to order kits.
  • Refer to Bulletin 6029 for information on normal levels of human cytokines detected with Bio-Plex cytokine kits, if desired.
  • Contact Bio-Plex core (orschlab@iupui.edu), Bio-Rad (Chris_Rutherford@bio-rad.com) and/or
    Millipore (steve.wowk@merckgroup.com) for assistance with sample collection protocols, experimental design, kit selection, or assistance with custom kits, as needed. Note: For planning purposes, each kit contains one 96-well plate capable of analyzing 78 samples or 39 if run in duplicate. Standards and blanks (run in duplicate) occupy 18 wells of each plate.
  • Order kit (avoid shipping over weekends), collect samples and store samples appropriately.
  • Complete the requisition form and submit to the Bio-Plex core. Bio-Plex core will contact you to schedule your assay time.
  • On analysis day, Bio-Plex core will contact you to inform that the Bio-Plex has passed calibration. At that time, thaw samples and dilute, if needed, using diluent provided in the kit and/or according to instructions in “Procedures”.
  • Deliver diluted samples to Bio-Plex core for analyses.

Please note: If you need to cancel or postpone your assay time, you must call the Bio-Plex core at (317) 278-2485 by 8 a.m. on assay day or risk being charged for instrument set-up.

Fees

Fees/Plate (effective July 1, 2014)

Internal IU faculty rates

External rates

$300

$495

Confidentiality

All data generated by the BioPlex core are kept confidential and will not be shared unless the owner provides specific written direction.

Prioritization of work

Work is prioritized based on a “first come, first served” policy.

Publication and authorship expectations

Authorship is not expected for routine operation of the BioPlex. If effort from core personnel exceeds routine operation of the BioPlex, authorship will be discussed and agreed upon prior to running the users’ samples.

Procedures
Preparation of blood samples

Note: Serum is strongly recommended over plasma. Accurate analyses of some targets cannot be ensured if using plasma. Also, minimize hemolysis as much as possible during processing as iron can interfere with some analyses.

  1. For serum, collect blood without anti-coagulant and allow to clot at room temperature for 30 minutes to 1 hour.
  2. If collecting plasma, use EDTA or sodium citrate as the anti-coagulant. DO NOT use heparin as the anti-coagulant as heparin has been reported to bind certain cytokines, interfering with detection.
  3. Centrifuge sample at 3000 rpm, 10 min, no brake, 4ºC.
  4. Store sample in aliquots of 50-100µL each (minimum of 20µL for single analysis or 40µL for duplicate analyses) at -70ºC until ready to analyze.
  5. On the day of analysis, when instructed by core, thaw sample and dilute. The recommended serum dilution for BioRad kits is 1:4, but user should use the dilution recommended for their particular kit. Dilutions should be made using sample diluent (provided in kit) to achieve a final volume of at least 75µl if analyzing one well per sample or 150µl if analyzing in duplicate.
  6. It is the user's responsibility to ensure that samples are free of particulates. Please remove all particulates by centrifugation or other suitable means before delivering to the core. We recommend performing a centrifugation step at 13,000 RPM on a benchtop microcentrifuge for 5-10 minutes.
  7. Keep diluted sample on ice and deliver to the Bio-Plex core.
Preparation of cell culture supernatant samples
  1. Centrifuge sample at 3000 rpm, 10 min, 4ºC. Collect supernatant.
  2. Store sample in aliquots of 75µL each (for single analysis) or 150µL (for duplicate analyses) at -70ºC until ready to analyze.
  3. On the day of analysis, when instructed by core, thaw samples and deliver on ice to the Bio-Plex core, along with tissue culture medium (~1ml) for dilution of standards. NOTE: There is usually no need to dilute tissue culture samples, but if dilution is necessary, dilute with tissue culture media to achieve a final volume of at least 75 µl if analyzing one well per sample or 150 µl if analyzing in duplicate.
  4. It is the user's responsibility to ensure that samples are free of particulates. Please remove all particulates by centrifugation or other suitable means before delivering to the core.
Preparation of cell lysate samples

Protocols for preparation of cell lysates from specific tissues are numerous and can be found in the links below, among others. This list is not meant to be inclusive and the investigator is encouraged to use any protocol he or she feels is best for their particular application. A tissue lysate protocol is also included in the BioRad kit for phosphoproteins, which may also be used for cytokine analyses of tissue lysates.

Tissue Cytokine Assays (refs)

Bio-Plex Article

Bio-Plex Cytokine