Please acknowledge the Diabetes Immune Monitoring Core in all publications resulting from full or partial use of our services, as this supports future Diabetes Research Center grant renewals.
“Data was obtained through the Diabetes Immune Monitoring Core of the Stanford Diabetes Research Center, supported by the NIH/NIDDK under Award Number NIH P30 DK116074.”
DIMC provides standardized, state-of-the-art immune monitoring assays at the RNA, protein, and cellular level, as well as archiving, reporting, and data mining support for clinical and translational studies. In partnership with the research community, we also work to test and develop new technologies for immune monitoring.
Immunoassays
We have standardized immunoassays for human and mouse cytokines, which can be used to assay serum, plasma, cell culture supernatants, tissue homogenates, or other fluids. We measure multiple cytokines simultaneously using either Luminex (up to 63 analytes) or MesoScale Discovery (MSD, up to 10 analytes) systems. We also provide assays for other soluble proteins on the MSD platform (see their website at www.mesoscale.com for available kits).
HIMC now offers services on the Olink platform
Olink proteomic panels achieve a high level of multiplexing with the use of Proximity Extension Assay Technology (PEA). Each biomarker is addressed by a matched pair of antibodies, coupled to unique, partially complementary oligonucleotides, and measured by quantitative real-time PCR. This dual recognition, DNA-coupled method provides readout specificity, avoiding antibody crossreactivity that may occur during the multiplexed assay process.
The Olink portfolio includes 12 human panels and one mouse panel each with 92 biomarkers per panel. https://www.olink.com/products/
The volume of sample required is extremely low however we request a minimum of 25ul of non thawed sample.
A few of the Metabolic panels require special collection tubes containing protease inhibitors.
The cost for each array is 10K and includes both materials and technical labor. An array can fit up to 84 samples run as singlet or 42 samples run in duplicate.
The most popular is the Inflammation panel: https://www.olink.com/products/inflammation/
DIMC will sponsor up to 10% of the cost. For more information or to discuss services please contact:
yaelhr@stanford.edu
Yael Rosenberg-Hasson,
Immunoassay & Technical Director
Human Immune Monitoring Center
1651 Page Mill Rd.Room 0280
Palo Alto CA 94304-5518
(650) 723-4984
Mass Cytometry
Mass cytometry, or CyTOF (DVS Sciences), is a variation of flow cytometry in which antibodies are labeled with heavy metal ion tags rather than fluorochromes. Readout is by time-of-flight mass spectrometry. This allows for the combination of many more antibody specificities in a single samples, without significant spillover between channels. We have standardized an immunophenotyping CyTOF panel with 23 antibodies, with other panels to come shortly. Experienced CyTOF users can also schedule time on the instrument to run independent experiments.
Genomics
The HIMC offers RNA and DNA extraction, Bioanalyzer services, gene expression and miRNA microarrays, and microfluidic qPCR arrays for human and mouse.
Flow Cytometry
Flow cytometry is used to assess immune cell phenotypes and functions. We offer standardized immunophenotyping, phosphoepitope, and intracellular cytokine assays for human blood cells. Our assays have all been optimized for use with cryopreserved PBMC; use with fresh samples must be coordinated in advance.
Nanoimmunoassays
The NanoPro nanoimmunoassay platform from Protein Simple is an automated microcapillary isoelectric focusing system. Once focused, proteins are crosslinked to the capillary wall, and subject to antibody probes, much like a Western blot. This platform, formerly in HIMC, is now offered through the Translational Applications Service Center (TASC). Contact Joanna Lilliental (jlili@stanford.edu) for more information.