Protein Profiling Services
Multiplex, ELISAs, & Enzymatic Assays

 Click the image above to learn more about the    Luminex xMAP technology.
ORB offers accurate quantification of cytokines, chemokines, phosphorylated proteins, MMPs, growth factors, and receptors using Luminex xMAP (multiple-analyte profiling) technology-based assays. To learn more about the Luminex technology click here. Many sample types are compatible with Luminex-based immunoassays including serum, plasma, tissue culture supernatants, cell lysates, urine, milk and even cerebrospinal fluid. The Luminex technology enables customizable protein assays that can measure up to 60 analytes simultaneously per sample. Generally, these assays have a dynamic range of 1,000—10,000 fold and sensitivity as low as 0.1pg/mL. Luminex xMAP microspheres can be generated in high volume single lots to facilitate reproducible results from high throughput screening of hundreds of samples.
ORB provides an end-to-end solution for protein profiling which includes panel selection, protein extraction, and data analysis. ORB's protein profiling services provide the highest quality data utilizing a wide selection of multi- and single-plex panels with analytes that are relevant to many diseases.  Kits run by ORB must be calibrated to NIBSC standards, and wherever possible, ORB’s protein profiling service utilizes multiplexing to create cost-competitive packages. Enzymatic and colorimetric assays can also be performed when needed.

Protein extraction services are available from a wide variety of tissues, and a modified Lowry assay is used to measure protein concentrations and normalize, if necessary. To optimize results, pilot studies can be conducted to determine appropriate sample dilutions prior to analyte characterization.

"ORB-GLP-certified-immunoassays"

  • Multiplex Protein Profiling
  • ELISAs
  • Colorimetric Assays
Learn more about ORB’s QA/QC Practices.
Each assay includes duplicate determination for each sample, a 7-9-point standard curve, and positive controls. ORB’s standard analysis packages include complimentary analysis using Luminex xPONENT software, calculated analyte concentrations, statistical analysis, graphical representations of detected analytes, and ANOVA and/or t-tests. Optional GLP certification is available for projects requiring documentation for subsequent patent or FDA applications.
  See our Price Sheet for the most up to date discounts on ORB’s protein profiling services.

Available Assays - Analyte Coverage

ORB sources assays from leading manufacturers such as Millipore, Life Technologies, and R&D Systems. See the bullet list below for a sample of the panels that are available, and use the custom panel tool below for a detailed view of available panels and the analyte coverage for each.
 
Millipore
  • Human, Mouse, and Rat Cytokines and Chemokines Over 75 human, 50 mouse, and 27 rat analytes. ORB runs up to 60-plex assays.
  • Human and Mouse Cytokine Receptors
  • Cancer Angiogenesis, circulating tumor markers, metastasis.
  • Endocrine Panels Human, Rat,and Canine panels for pancreatic, pituitary and brain peptide hormones.
  • Metabolic Panels Human, Mouse, Rat adipokines and markers of diabetes and adipocyte cells.
  • Gut Hormones Human, Mouse, Rat, Canine.
  • Cardiovascular Disease Panels Human, Mouse, and Rat markers of cardiac damage, apolipoproteins, clotting factors, and serum proteins.
  • Gut Hormones Human, Mouse, Rat, Canine.
R & D Systems
  • Human Obesity Panel Up to 10-plex
  • Human Adhesion Molecule 4-Plex
  • Human Angiogenesis Panel Up to 11 angiogenesis markers.
  • Human Matrix Metalloproteinase 8-Plex.
  • Additional Panels for Skin, Bone, and Neuronal Markers available.

Download ORB’s price sheet to obtain multi- and single-plex panel pricing. Contact Us if your plex of interest is not shown.
 

Custom Panel Selection Tool - Design and Submit Requests for Custom Luminex Assays

Click on the species button below to select your analytes and design your custom assay using our custom panel selection tool. We offer hundreds of panels for Human, Mouse, Rat, and other species using multiplex bead-based Luminex® kits from Millipore, Life Technologies and R&D systems. Contact us for other species.


Sample Submission Instructions

Volume requirements for protein profiling services are assay specific. Contact us for more information or if your sample type is not listed below. A minimum of 5 ml of unconditioned media is needed, for standard and control dilution, in studies analyzing cell culture media. For projects examining analyte levels in cells or tissue, protein extracts must usually contain a minimum of 3 micrograms of protein per microliter in order to obtain acceptable results; if submitting these sample types please use ORB's homogenization protocol(s) linked in the Table below. Clients may also submit cell pellets or tissues for homogenization at ORB.

Sample Type Sample Preparation Protocol Sample Submission Checklist
Cells Cell culture Cell culture
Tissues Tissues Tissues
Biofluids See sample submission checklist Biofluids


ORB Protein Profiling Experience

Inflammatory Bowel Disease

Cytokines in inflammatory bowel disease
Figure 2. Depiction of cytokines which play essential roles in the pathogenesis of IBD (Neurath).

A collaboration between the Medical College of Wisconsin and the Mayo Clinic sought to better define the pathological mechanisms, specifically the immune state, that regulating Crohn’s disease and ulcerative colitis, two major types of inflammatory bowel disease (IBD). This team utilized ORB’s multiplex protein profiling service to measure the levels of pro-inflammatory (TNF-α, IL-1α, IL-1β, IL-2, IL-6, IL-8 and IL-17α) and anti-inflammatory (TGF-β1, TGF-β2, TGF-β3, IL-10, IL-1RN) cytokines as well as soluble receptors (TNF-R1 and TNF-2) in human plasma samples. The researchers identified a TGF-β and IL-10 dependent immuoregulatory signature which may be utilized as a basis for additional analyses to develop a set of predictive biomarkers for IBD patients1.


Respiratory Illness

Protein phosphatase 2A (PP2A) is serine/ threonine phosphatase that is essential for a wide variety of cellular processes such as DNA replication, protein translation, cytoskeleton dynamics, and apoptosis. It is considered a tumor suppressor gene because blocking PP2A activity in cells can promote transformation and also various mutations in the PP2A gene have been identified in tumors3. Wallace and co-workers observed increased PP2A expression in the lungs of mice and human airway epithelial cells exposed to cigarette smoke. The researchers contracted with ORB to perform multiplex protein profiling of matrix metalloproteinases and interleukin-8 in small airway epithelial cells after manipulation of PP2A levels and exposure to cigarette smoke4.  The results of these assays showed that transfected PP2A could block the increase in levels of MMP1 and MMP-3 in conditioned media in response to cigarette smoke exposure, leading to the hypothesis that PP2A may be a potential therapeutic target for chronic obstructive pulmonary disease5.

Oncology

An investigation of the differences between biomarker signatures of low and high risk neuroblastoma patients employed ORB’s multiplex protein profiling service to quantitate IL-1β, IL-2, IL-4, IL-6, IL-10, IL-17α, IFN-γ, TNF-α, TGF-β, MCP-1, GM-CSF and RANTES in serum samples. From this work, researchers determined low risk patients showed higher levels of cytokines participating in innate immune responses whereas high risk patients show skewed adaptive responses6.

Contact Us to discuss how ORB’s protein profiling services can advance your research!

 
References
  1. Gurram, B., Salzman, N. H., Kaldunski, M. L., Jia, S., Li, B. U. K., Stephens, M., & Hessner, M. J. (2016). Plasma‐induced signatures reveal an extracellular milieu possessing an immunoregulatory bias in treatment‐naive paediatric inflammatory bowel disease. Clinical & Experimental Immunology184(1), 36-49.
  2. Neurath, M. F. (2014). Cytokines in inflammatory bowel disease. Nature Reviews Immunology14(5), 329-342.
  3. Seshacharyulu, P., Pandey, P., Datta, K., & Batra, S. K. (2013). Phosphatase: PP2A structural importance, regulation and its aberrant expression in cancer. Cancer letters335(1), 9-18.
  4. Wallace, A. M., Hardigan, A., Geraghty, P., Salim, S., Gaffney, A., Thankachen, J., ... & Foronjy, R. F. (2012). Protein Phosphatase 2a (Pp2a) Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung. Toxicological Sciences, kfr351.
  5. Wallace, A. M., Hardigan, A., Geraghty, P., Salim, S., Gaffney, A., Thankachen, J., ... & Foronjy, R. F. (2012). Protein Phosphatase 2a (Pp2a) Regulates Innate Immune and Proteolytic Responses to Cigarette Smoke Exposure in the Lung. Toxicological Sciences, kfr351.
  6. Gowda, M., Godder, K., Kmieciak, M., Worschech, A., Ascierto, M. L., Wang, E., & Manjili, M. H. (2011). Distinct signatures of the immune responses in low risk versus high risk neuroblastoma. Journal of translational medicine9(1), 170.
 
 
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