Getting Started

Preparing Your Samples for Submission

1. Design your experiment based on your biological question. If your data needs to be analyzed statistically, please discuss your experimental design with your bioinformatics group prior to setting up your experiment. (MD Anderson users consult the Bioinformatics Shared Resources group)
   
   
2. Prepare your RPPA samples (protein lysates) from cultured cells or tissue samples based on the procedures provided on the Resources page. 
   • A minimum of 80 μl of protein lysate per sample is required for RPPA analysis.
      
3. Adjust the protein concentration to approximately 1.5 μg/μl.
   
   
4. Denature the protein by adding 4× SDS + 2-mercaptoethanol. (Final concentration after denaturation should be ~1 μg/μl).
   • For quality purposes, we recommend a final concentration of no lower than 0.5 µg/µl for cultured cells and 0.75 µg/µl for tissues.
      
5. If you choose to submit cell pellets or tissue for the protein extraction to be performed at our Core Facility, we require a cell pellet with a minimum of 1 million-5 million cells depending on the cell size (about a 5 µl pellet) or a piece of tissue with a minimum of 15 mg (about the size of a large grain of rice) with at least 60% cellularity. 

Current Sample Types Accepted for RPPA 

• Cultured Cells
• Human Tissue
• Mouse Tissue
• Mouse Xenografts

We currently do not accept FFPE samples for RPPA processing, due to the lack of quality and analyzable protein present within this sample type. 

Biological replicates are encouraged, but not required. (See Frequently Asked Questions for more information about replicates.) 

• Please note that each replicate submitted will be considered as a separate sample.
  
  
• If you submit replicates as separate samples, please create a unique name for each sample and submit each replicate in a separate vial. 
  • Unique names can be created by tagging the sample name with e.g., “-1, -2, -3.”
  • Example: Sample A-1, Sample A-2, Sample A-3.     

Submitting Your Samples to the RPPA Core

1. Sign up for an iLab Solutions account by clicking on or copying and pasting the following web address in your browser: https://mdanderson.ilabsolutions.com.
   
2. After your account is set up, request services through the MDACC RPPA-Functional Proteomics iLab Page.
   
3. After your online request is fully approved, you will be notified by email either to schedule sample drop off or shipment and instructions. 
   • Note that the submitter AND PI or Financial Manager of the iLab request must both log into their own iLab account and agree to the request before it is considered fully approved.
4. Label the tops of the sample vials (1.5 ml microcentrifuge tubes or 2 ml Precellys tubes) with the sample number (not the sample name) in the order listed in your iLab sample list.
   • Do not put labels or tape around the sides of the sample vials as these must be removed before processing. 
5. Pack samples on dry ice.
   
6. Deliver/Ship to the RPPA Core after you coordinate a drop-off/shipment date by email. 
   • MD Anderson submitters (after iLab approval by the Core) may fill out a Courier Request Form to drop off samples to the Zayed building. 

Our goal is to deliver results within 8-10 weeks after receiving samples. However, based on sample collection, workload, equipment failure or software issues, processing and analysis may occasionally take up to 12 weeks. We do everything possible to shorten the timeline while maintaining high quality data at a low cost. 

Payment 

Inside MD Anderson: PeopleSoft Account Number

Prior to requesting services in iLab, please do the following:

• Contact your Financial Manager to enable access to your funding source(s)
• Set up your funding source(s) within your iLab account 

Outside MD Anderson: Purchase Order (PO) or Credit Card 

An electronic (pdf file) copy of the PO or Credit Card Charge Authorization Form must be either emailed to ccsgrppa@mdanderson.org or attached to the iLab request before you can submit your samples. The Core will not accept your samples without this.