Overview
The Nanoscale Protein Analysis Section of the CCR Collaborative Protein Technology Resource (CPTR) provides expertise and service in state-of-the-art protein analysis technologies to advance CCR research in basic discovery and translation/clinical studies. Applications include:
- Quantitative profiling of cell signaling and post-translational modification
- Measurement of cytokine, metabolite and serum/plasma markers
- Identification and validation of biomarkers & therapeutic targets
- Assessment of on‐ and off‐target drug activity and pharmacodynamic evaluation
- Study of tumor microenvironment and single-cell signaling
Established Technologies
Automated capillary immunoassay system – Simple Western technology
- High-resolution MW (size-based) or isoelectric-focusing (IEF, charge-based) separation, followed by target-specific immunoprobing to profile proteins and respective post-translational modification isoforms
- Nanogram level of protein analysis for extremely small and precious samples, e.g. stem cells, primary cells, micro-dissected samples, fine needle aspirates, non- or minimally- invasive collected patient specimens etc.
- Provides precise and accurate measurement of protein expression level and activation status, highly quantitative data, good assay sensitivity, dynamic range and reproducibility
- About three hundred assays covering key signaling pathways have been established and successfully applied for functional proteomic research, will validate assays for other targets as needed
- Assay readiness for bench to bed transfer
In-solution multiplex ELISA assays – Luminex xMAP Technology
- Up to 50-plex simultaneous protein target analysis with small sample consumption
- Analysis of cytokine, metabolite, immune response, serum/plasma biomarkers etc.
- Clinical applicable assay performance
Immunofluorescence imaging platform – CODEX (CO-Detection by indEXing) technology
- Provides full spatial context and is not limited to just regions of interest (ROI)
- Provides single-cell resolution down to 600 nm or 250 nm depending on microscope objective used (20X and 40X respectively)
- Single-step staining and gentle fluorophore removal preserves the sample for ROI analysis
- Simple, benchtop fluidics system that is cost-effective and simple to implement in any research lab
Developing Technologies
Single-cell Western system
- Western-based protein analysis on 1000-2000 single-cells in parallel
- Measures proteins hard-to-detect by FACS, such as isoforms, phosphorylated proteins, transcription factors, intracellular proteins etc.
- Single cell signaling profiling and population heterogeneity study
We actively evaluate, develop, acquire and implement innovative protein analysis technologies to accommodate the demands from the CCR/NCI/NIH community.
Digital Spatial Profiling
- NanoString GeoMx Digital Spatial Profiling (GeoMx DSP)
- Offered in collaboration with the CCR Genomics Core
- FFPE or fresh frozen
- Profiling data for 100s of validated proteins
- Cancer Trascriptome Atlas (CTA) > 1,800 gene
- Whole Transcriptome Atlas (WTA) > 18,000 genes
User Guidelines
- Projects are collaborative and subject to the approval of resource advisory committee.
- Investigators are expected to share the cost of certain consumable reagents, for which CCR investigators are eligible for a 50% subsidy from the CCR Office of Science and Technology Resource (OSTR).
- Staff are available to assist on project design, data interpretation and study milestone evaluation, as well as general consultation on proteomic analysis questions.
Keywords
4E-BP1ABCG2AKTALAS1AMPKBIMBLMBmi1CIAPCODEXCPTRCaspaseChk1CofilinCyclinDDB1DNA damageDNMT1DUSP3E2F-1EGFREGR1ELISAERB2ERKFADDFHL1FHL2FLIPFlagFosBFox03AFra1GAB1GAPDHGSTGlucose-6-phosphate dehydrogenase (G6PD)HSP70HSP70sIntegrinIκBαIκκαJAK2JNKJunBKIDLIMK2LuminexMAGPIXMAP kinaseMCM2MCM4MCM5MEKMiloMultiplex Tissue ImagingNFκBNFκB p65NFκB1 p105/p50NFκB2 p100/p52NanoProTMPARPPI3 Kinase p110 α/βPI3 kinasePKCPLCγ1PLK1PTENPeggy SuePeggy SueTMPeggyTMProteinSimpleProteinsProteomicsRIPROCKRafRasGRPRbRelBS6 Ribosomal proteinSHC1SMAC/DiabloSMADSMARCB1SRCSTAG2STATSallyTMSimple WesternSpatial TranscriptomicsTACC3TRAF1Thioredoxin 1VinculinWesTMXIAPanalyteapoptosisbiomarkerc-Junc-Mycc-RelcFoscapillary immunoassaycell cyclecell signalingchemokineclinical trialcore biopsycultured mammalian cellscytokinedrug developmentepigeneticsProteins and ProteomicsSingle Cell Analysisfine needle aspiratesheterogeneityimmunoassayinflammationloading controlsmTORmicro-dissectionmolecular profilingmultiplex ELISAmultiplex immunoassaynano-fluidic immunoassaynci-corep-glycoproteinp53p70 S6peripheral blood mononuclear cells (PMBCs)pharmcodynamic analysisphospho-Histone H2A.X (S139)phosphorylation; post-translational modificationplasma cellspreclinic and clinic sample analysisproliferationprotein characterizationprotein complexprotein quantificationsingle cellsubcellular fractionationtarget therapytissue samplestranscription factortyrosine kinasewesternwestern blotxMAPα-Tubulinβ-Actinβ-Catenin
Contact Information
Name:
Noemi Kedei, MD
Title:
Facility Head
Address:
Building 41, Room D310Bethesda, MD 20892
Phone:
E-Mail:
Additional Core Staff
Name:
Maria O. Hernandez, PhD
Title:
Scientist
Phone:
E-Mail: