Shared Facilities

Bio-Plex

Policies & Procedures

To use the core, please do the following:

  • Set up an account for invoicing.
    • Complete the “Authorization form.”
    • Email the completed form to Kristina Taylor (kristtay@iu.edu).
  • Find an “off-the-shelf” kit or create a “custom” kit that will identify your specific targets, tissue, and species. Some resources:
    • xMAP kit finder (Select Luminex 100/200 instrument)
    • Assay Builder Web site for Bio-Rad bio-plex products
    • Published literature
    • Technical support from your favorite company (ex. Millipore: Jason.Lemberg@emdmillipore.com, or Bio-Rad: Chris_Rutherford@bio-rad.com)
  • Plan your experiment.
    • Each kit contains enough reagents for 96 reactions. Standards and blanks consume 18-20 reactions, leaving enough reagents for 76-78 samples run in singlet, or 38-39 samples run in duplicate, as you prefer.           
    • Collect samples following information in the “Technical Procedures” section below.
    • Additional instructions on sample preparation are included in kits, on the Bio-Rad website (keyword: “Bio-Plex”), and on the Millipore website (keyword “Milliplex”).
  • Schedule time with the Multiplex core.
    • Complete the requisition form & submit to the Multiplex core (orschlab@iupui.edu).
    • The Multiplex core will contact you to schedule your assay time.
    • On the day of analysis, core personnel will contact you when the Bio-Plex instrument has passed calibration. At that time, thaw and deliver your samples (on ice) to the core.
    • If you need to cancel or postpone your assay time, you must call the Multiplex core at (317) 278-2485 by 8 a.m. on analysis day or risk being charged for instrument set-up.
Fees

Fees/Plate (effective July 1, 2015)

Internal IU faculty rates

External rates

$300

$495

Confidentiality

All data generated by the Multiplex 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 Multiplex. If effort from core personnel exceeds routine operation of the Multiplex, authorship will be discussed and agreed upon prior to running the users’ samples.

TechnicalProcedures
Preparation of blood samples
  1. Serum is strongly recommended over plasma. If collecting plasma, use EDTA or sodium citrate as the anticoagulant. Heparin may interfere with detection of some cytokines, and is thus NOT recommended as an anticoagulant.
  2. Minimize hemolysis as much as possible as iron may interfere with some analyses.
  3. Store samples in aliquots of 50-100µL each (minimum of 20µL for singlet analysis or 40µL for duplicate analyses) at -70ºC.
  4. On the day of analysis, when instructed by core, thaw samples and dilute using sample diluent & recommended dilutions (provided in kit) to achieve a final volume of at least 75µl if analyzing in singlet or 150µl if analyzing in duplicate.
  5. It is the user's responsibility to ensure that samples are free of particulates by centrifugation (we recommend 13,000 RPM on a benchtop microcentrifuge for 5-10 minutes) or other suitable means before delivering to the core on ice.
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 singlet analysis) or 150µL (for duplicate analyses) at -70ºC.
  3. On the day of analysis, when instructed by core, thaw samples and deliver on ice to the Multiplex 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 in singlet or 150 µl if analyzing in duplicate.
  4. It is the user's responsibility to ensure that samples are free of particulates by centrifugation (we recommend 13,000 RPM on a benchtop microcentrifuge for 5-10 minutes) or other suitable means before delivering to the core on ice.
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 Bio-Rad kit for phosphoproteins, which may also be used for cytokine analyses of tissue lysates.

Bulletin 6029 (Rename as: Normal Physiological Levels of Human Cytokines Using the Bio-Plex ProTM Cytokine Assays)

Tissue Cytokine Assays (refs) (Rename as: Bio-PlexTM Cytokine Assay References)

Bio-Plex Article (Rename as: Optimization of multiplexed bead-based cytokine immunoassays for rat serum and brain tissue. Hulse et al., 2004)

Bio-Plex Cytokine (Rename as: Use of the Bio-Plex cytokine immunoassay to determine cytokine expression levels in the intestinal mucosa)