The Immunology Core provides a wide range of services and reagents to benefit the immunological research of the Penn CFAR Community. Services are provided by facilities in both the University of Pennsylvania and the Children's Hospital of Philadelphia Research Institute and are housed at several sites. Services include:
- Primary Cell Purification (Riley Lab, 8th Floor Similow Center for Translational Research)
- Freshly isolated primary PBMCs, CD4, CD8, T cells, NK, monocytes and B cells
- CCR5 deficient T cells and monocytes from Δ32 donors
- Standard Immunological Assays (12th Floor Abramson Research Center, CHOP)
- Cell Based Assays
- Flow Based Assays and Services
- Cellular, Humoral and Molecular Immunology assays (HIC, 410 & 704 Stellar Chance)
- Specimen Processing, Banking and Sample Shipment
- Luminex Assays
- Flow Cytometry
- Sequencing of Human and Murine IgH rearrangement
- Sequencing of Human TCR Vβ Rearrangements
- Amplification and cloning of Ig rearrangements
- Data analysis pipeline for repertoire studies
- Customized data analysis
- Scientific Consultation and Assay Development
- BSL-3 Cell Sorting Facility
- Small Animal Model of HIV-1 Infection
Standard Immunological Assays
Cell Based Assays
Contact Nancy Tustin at firstname.lastname@example.org or 215-590-2043 for pricing and scheduling.
- Functional Natural Killer Cell Assay (FNKA)--The lab is CLIA certified for this assay, so the results of the assay can be used for patient management. The functional capability of NK cells is determined in vitro by exposing them to chromium labeled NK sensitive target cells. NK cell activity is quantified in this assay by the measurement of chromium released from NK cell lysed targets. This lytic activity is normalized to reflect the number of peripheral blood mononuclear cells and natural killer cells per 107 cells required to lyse 20% of the chromium-labeled targets. This is referred to as the LU20 PBMC and LU20 NK, respectively.
- Cytokine assays on cell culture supernatants and biological fluids by ELISA. Pricing is calculated as the cost of the ELISA kit plus labor.
- Lymphoproliferative assays to phytohemagglutinin (PHA), pokeweed mitogen (PWM) and Candida
- Other assays and services are available upon request (separation of PBMC from whole blood, separation and cryopreservation of plasma, Luminex, etc)
Flow Based Assays and Services
Contact Florin Tuluc at email@example.com or 267-426-5350 for pricing and scheduling.
Sample processing (surface and intracellular staining), functional assays, analyses, and access to flow cytometry analyzers are offered on a fee-for-service basis. Most common assays include immunophenotyping of human blood samples from clinical studies:
- Flow cytometric, single platform CD4 counts (CLIA and IQA certified)
- Custom multi-color immunophenotyping of leukocyte subpopulations :
- leukocyte subsets (T, B, NK cells, monocytes)
- T cells subsets (naive, memory)
- immunoregulatory cells (Tregs)
- NK cells: expression of inhibitory/activating NK receptors
- Circulating stem cells and progenitors
- Intracellular cytokine panels
- Custom panels of antibodies
The laboratory routinely performs full service testing on whole blood as well as fresh and cryopreserved PBMCs. Custom panels of antibodies are available upon request.
Functional flow cytometry and bead-based assays are also available as full-service to our users:
- Cell proliferation and viability
- Cell cycle
- Mitochondria membrane potential and generation of reactive oxygen species; mitochondrial mass
- Intracellular calcium assays by flow cytometry
- Protein phosphorylation using phospho-specific antibodies
- Cytometric Bead Array for TH1/TH2 Cytokines
- Cytometric Bead Array Flex Human or Mouse custom cytokine assays
- Other flow cytometry assays are available upon request.
Four analyzers are available in our facility and they may be operated by trained users:
- LSRFortessa (blue, red, yellow-green, violet and UV lasers; 18 color detectors).
- LSRII (blue, red, and UV lasers; 8 color detectors)
- FACSCalibur (blue and red lasers; 4 color detectors)
- Accuri C6 (blue and red lasers; 4 color detectors)
The laboratory currently operates a MoFlo XDP cell sorter (Beckman-Coulter) encased in a Class II biosafety cabinet.
Unasisted use of analyzers and cell sorting services can be scheduled online by registered users at http://www.research.chop.edu/cores/flowcytometry/skedit