Multiplex Analysis Core

Policies & Procedures

GETTING STARTED

To use the core, please do the following: 

Set up an account for invoicing

  1. Complete the “Authorization form.”
  2. E-mail the completed form to Kristina Taylor (kristtay@iu.edu) and the MAC at pplett@iu.edu.

Find an “off-the-shelf” kit or create a “custom” kit that will identify your specific targets, tissue, and species. Some resources:

  1. xMAP kit finder (Select Luminex 100/200 instrument)
  2. Assay Builder Web site for Bio-Rad bio-plex products
  3. Published literature
  4. Technical support from your favorite company (see contacts)

Plan your experiment

  1. 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.           
  2. Collect samples following information in the “Technical Procedures” section below.
  3. Additional instructions on sample preparation are included in kits, or on the vendor websites (see contacts). 

Schedule time with the MAC

  1. Request the requisition form from the MAC (pplett@iu.edu or hfeng@iu.edu).
  2. The MAC will contact you to schedule your assay time.
  3. Submit the requisition form at least 24 hours before your scheduled run.
  4. If you need to cancel or postpone your assay time the day of the run, please call (317) 278-2485 the MAC by 9 a.m. on the day of analysis or risk being charged for instrument set-up.
Fees

Number of  analytes Price / Plate - Internal Price / Plate - External
1-5 $261 $431
6-10 $273 $450
11-15 $285 $470
16-20 $297 $490
21-25 $309 $510
26-30 $321 $530
31-35 $333 $549
36-40 $345 $569
41-45 $357 $589
46-50 $369 $609
51-55 $381 $629
56-60 $393 $648
61-65 $405 $668
Add $10 / plate for kits requiring a dilution step.

The IU Simon Cancer Center will invoice the investigator when the work in completed using the account number provided on the Authorization form.

Policies
Confidentiality

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

Cost Recovery/Payment

Payment is expected for services rendered. An IU account number or a purchase order number (PO#) must be provided before service is provided, and the PI or person responsible for payment must sign and date the Authorization form. Full payment is expected if data are obtained regardless whether or not it represents the result expected by the investigator. Partial or waved payments may be accepted in cases of instrument malfunction. The MAC does not provide sample storage after analyses. Samples not retrieved from the MAC within one week of analyses will be discarded.

Prioritization of work

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

Acknowledgement, publication and authorship expectations

Authorship is not expected when routine analyses are performed by the Multiplex Analysis Core. However, if effort from core personnel exceeds that of routine analyses, authorship will be discussed and agreed upon prior to running the users’ samples. If research supported by the MAC results in publication, please acknowledge this support by including the following in your publication(s): We thank the Multiplex Analysis Core at the Indiana University Melvin and Bren Simon Cancer Center for providing support in analyzing samples and interpretation of data. 

Please notify the MAC of your publication by e-mailing us at corschel@iu.edu or pplett@iu.edu

Conflict Resolution

The Director will discuss all issues with the investigator in order to resolve them. Should issues (such as charge disputes, authorship, and conflict of interest) arise that cannot be resolved in this manner, the Advisory Committee will assist in conflict resolution.

Submitting Material in Compliance with Regulatory Requirements

Material or health-related information submitted to a Core for analysis must be obtained in compliance with the most current regulatory and legal requirements (including material transfer agreements). It is the responsibility of the investigator to make sure such requirements are fulfilled.  It is also the responsibility of the investigator to comply with all safety standards (e.g. Radiation and Bio-Safety) and fully disclose the potential hazards of all materials submitted to a Core.

Technical Procedures

Preparation of blood samples

  1. Serum is recommended over plasma, but either will be ok. When heparin is used as the anticoagulant in plasma samples, please check your kit manual for the proper heparin concentration (Millipore suggests to use <10 IU/mL Heparin in the plasma).
  2. Minimize hemolysis as much as possible as iron may interfere with some analyses.
  3. Store samples in small aliquots to avoid freeze thawing if running multiple assays. Store approximately 50ul per single well in the Bioplex assay, or ~100µL if duplicate wells will be run. Note that the volume required will depend on which analyte is assayed, as some require dilution before being assayed.
  4. On your scheduled analysis day, bring your samples on dry ice and kits on wet ice to the MAC.
  5. To ensure that the samples are free of particulates, we will centrifuge the samples at 13,000 RPM on a benchtop microcentrifuge for 10 minutes.

Preparation of cell culture supernatant samples

  1. Centrifuge samples at 3000 RPM for 10 min at 4ºC. Harvest supernatant.
  2. Store samples in aliquots of 75µL each (for singlet analysis) or 150µL (for duplicate analyses) at -70ºC.
  3. On your scheduled analysis day, thaw samples and deliver them to the MAC on ice, 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. To ensure that the samples are free of particulates, we will centrifuge the samples at 13,000 RPM on a benchtop microcentrifuge for 10 minutes.

Preparation of cell lysate samples

Protocols for preparation of cell lysates from specific tissues are numerous and can be found on vendor websites and in publications. 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. 

Additional sample preparation protocols can be found on the vendor websites.