Preparing Your Samples for Submission
- Design your experiment based on 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.
- Prepare your RPPA samples (protein lysates) from cultured cells or tissue samples based on the procedures provided on the Antibody Information and Protocols page.
- Adjust the protein concentration to approximately 1.5 µg/µl (no less than 0.75 µg/µl).
- Denature protein by 4x SDS + 2-mercaptoethanol. (Final concentration after denaturation should be ~1 µg/µl).
- A minimum 80 µl of protein lysate is required for RPPA analysis.
- 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 volume of about 5 µl (1 million-5 million cells depending on the cell type) or a piece of tissue about the size of a grain of rice (approx. 10 mg).
Current Sample Types Accepted for RPPA
- Cultured Cells
- Human Tissue
- Mouse Tissue
- Mouse Xenografts
We 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.
- If you choose to submit replicates, please label each one as a separate sample in a separate vial. Label on the Sample List with non-identical names (i.e. SampleA-1, SampleA-2, SampleA-3...).
Submitting Your Samples to the RPPA Core
- Sign-up for an iLab Solutions account by cutting and pasting the following web address in your browser: https://mdanderson.ilabsolutions.com.
- Once your account is set up, request services through the RPPA Core iLab Page.
- Once your online request is fully approved, you will be notified by email to set an appointment time for drop off/shipment instructions. (**Note that the submitter AND PI and/or Financial Manager of the iLab request will both have to log into their own iLab account and "agree" to the request before it is considered fully approved/completed.**)
- Label sample vials clearly on top in numerical order according to your sample list submitted online. (Do not put labels/tape around the outside of the vials).
- Pack samples on dry ice.
- Deliver/Ship to RPPA Core after you coordinate a drop-off/shipment date by email.