University of Pittsburgh Cancer Institute (UPCI)

CCSG Acknowledgement

Required CCSG Acknowledgement

The NCI requires that publications acknowledge the UPCI CCSG support, and they are tracking compliance. If a UPCI CCSG-supported Shared Resource provided data used in your publication, please include the following statement in the acknowledgment section of your publication(s):

"This project used the UPCI [insert name(s) of shared resource(s)] that [is/are] supported in part by award P30CA047904."

Shared Resource Directors: Please make sure to include this statement on all of your order forms, contracts, etc. as a reminder to your users to acknowledge the UPCI CCSG support.


CPF Luminex Core Laboratory

The versatile laboratory multianalyte profiling (LabMAP™) system developed by Luminex Corporation, located in Austin, Texas, is a flexible, easy-to-use protein analysis system that permits the simultaneous quantitative analysis of up to 100 different proteins, peptides, or DNA molecules in a single microtiter well. This multiplexing feature allows one to obtain a wealth of information using a minimum amount of sample. Multiplexing enables a comprehensive overview of complex biological systems and the inclusion of in-process controls. In addition, sample volume requirements are reduced and assay throughput is dramatically increased. Potential applications include:

  • Functional genomics
  • Validation of drug leads
  • Secondary screening
  • Absorption and distribution
  • Metabolism
  • High-throughput ELISA and Western blot assays

With the Luminex suspension array system, you can:

  • Quantitate up to 100 protein targets from culture media, sera, and other matrices simultaneously
  • Automatically analyze up to 96 samples in 60 minutes (general format: 88 sample wells + 8 standard wells)
  • Dramatically increase the amount of useful data

Luminex

Available Analytes

Commercial Kits

Core Developed Assays

Circulating Antigens:
Growth and Angiogenic Factors: EGFR, angiostatin, endostatin, thrombospondin, IL-18
Cancer Antigens: CA 125, CA 15-3, CEA, AFP, CA 19-9, CA72-4, mammaglobin
Growth Factors/Oncogenes: Bcl-2, EGFR, Her2/neu, IGFBP-1, IGFBP-2, SCC
Proteases: PSA, fPSA, Kallikreins 10, Transglutaminase II
Other Markers: CD105, Cyfra 21-1, EPCAM, HE4, mesothelin, MICA, NSE, Ostepontin, TTR, SAA, SMR, Hsp70, VWF, CEACAM-1, CEACAM-6, ALCAM, HIF-1alpha

Please contact the facility for specific kit information.

Selected Luminex Partners

Multiplexed Cell Signaling Protein Detection (Life Technologies)

Several companies offer kits for the multiplex detection for human, non-human primate, canine, porcine, mouse, and rat cytokines. They also offer a wide variety of ancillary and “general-purpose” reagents, including microspheres labeled with capture probes that specifically bind avidin (biotin), immunoglobulins (secondary antibodies and protein A/G), or common fusion protein tags (Ni2+ and glutathione).

For example, Life Technologies offers “AKT Pathway Total 7-Plex” and “AKT Pathway Phospho 7-Plex” platforms, which are specifically designed for quantifying the total proteins: IR, IGF-1R, IRS-1, Akt, PRAS40, p70S6K, and GSK-3β or alternatively the phosphorylated proteins: IR[pYpY1162/1163], IGF-1R[pYpY1135/1136], IRS-1[pS312], Akt[pS473], PRAS40[pT246], p70S6K[pTpS421/424], and GSK-3β[pS9].

Multiplexed Transcription Factors Technology (Active Motif)

Luminex® Transcription Factor Multiplex Assay schematicThe Multiplexed Transcription Factor Profiling System allows researchers to identify proteins that drive biological responses. It employs a powerful multiplexing approach for simultaneously measuring the binding of many different transcription factors to their unique regulatory DNA elements. The technology dramatically accelerates studies ranging from basic research in signal transduction and proteomics to target validation, compound screening, toxicity testing and disease characterization. This innovative technology offers unique benefits to researchers studying gene regulation. Unlike conventional methods such as EMSAs, each assay measures the presence of multiple transcription factors simultaneously, allowing powerful comparisons of different cell lines and treatments.

QuantiGene® Plex Assay (Affymetrix)

The QuantiGene Plex Assay is a hybridization-based assay using the xMAP® Luminex® magnetic beads and performed on 96-well plates. The assay is based on DIRECT quantification of the RNA targets using xMAP Luminex beads for multiplexing of 3 to 80 RNA targets and branched DNA (bDNA) signal amplification technology. On the first day, the sample is lysed to release the RNAs, and incubated overnight with target specific probe sets panel. On the second day, the signal amplification tree is built via sequential hybridization of PreAmplifier (PreAmp), Amplifier (Amp) and Label Probe (LP). Each amplification unit gives a 400 x signal amplification. and there are six amplification units per target RNA copy leaning to a 2,400 x signal amplification per copy RNA. The signal is detected by adding SAPE substrate and using a Luminex instrument for the read-out.

More information on the QuantiGene Assay is available here.