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Insights from Multiplex Assays: Small Samples, Big Impact!
Monitor Multiple Genes and Proteins with Minimal Sample.
Richer Gene and Protein Info from Fewer Cells!
Quantify any Proteins and those "Undetectable".
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Simple Western, Western Blot, In-Cell Western Services

Simple Western (Capillary-based Nano-Proteomic Immunoassay, Automatic & Quantitative Western Blot)  

  • (Courtesy of ProteinSimple)
    Quantitation of multiple target proteins / biomarkers in samples of limit amount, such as those from cells of live animals and patients including fine needle aspirates (FNA), laser capture microdissection (LCM), as well as serum, plasma, cell culture, etc
  • > 2000 antibodies have been validated for protein quantitation
  • Real-time toxicological and PK/PD monitoring drug candidates in experimental animals
  • Monitoring signaling transduction events in limited samples (as low as 25 cells), great for those from stem cells, bone marrow cells, sorted cells, etc
  • Discovery and validation of biomarkers to support developing predictive and companion diagnostics
  • Characterization of monoclonal and polyclonal antibody binding affinities
  • Characterization of specificity of antibody to the target protein of interest
  • Characterization of purity and charge heterogeneity of biopharmaceuticals
  • Phospho-protein profiling and quantitation of post-translational protein modifications using pan antibodies (after protein separation by charge/pI values) or using isoform specific antibodies (after protein separation by size/MW)
  • Screening test compounds for effects on expression or modifications of any protein or signaling molecule of interest

MW (Size)-Based SIMPLE WESTERN®


PI (IsoElectric Focusing)-BASED SIMPLE WESTERN®

        

Traditional Western Blot (Semi-Quantitative, with SDS-PAGE)  

  • Our traditional Western blot uses high quality pre-cast gels with various chemistries (such as SDS-PAGE, Native-PAGE, etc), as well as many well-validated specific antibodies, to obtain high resolution and consistent performance among multiple experiments.
  • LiCor Odyssey Near-Infrared Imaging technology is used to scan the probed target proteins giving a wider dynamic range, two-color (green and red) detection and quantification with higher sensitivity when compared common chemiluminescence methods.



In-Cell Western Blot (In-Cell ELISA™)  

  • Our assay combines the specificity of Western blot with the reproducibility and throughput of ELISA, is a simple and cost-effective means for quantification of intracellular signaling in whole cells.
  • This assay involves seeding cells in 96- or 384-well microtiter plates followed by fixation/permeabilization and subsequent labeling with activation state-specific or control antibodies, infrared-conjugated secondary antibodies, and/or far-red DNA dye.
  • The well-level data acquired by the Odyssey® Imaging System permits accurate measurement of signal induction or inhibition, with the potential for multiple treatments, end points, and replicates on each plate.
  • Our assays quickly, accurately measure relative protein levels in many samples, detect proteins in situ in a relevant cellular context.
  • This is used for high-through put analysis of: protein phosphorylation (effects of drug compounds on signaling pathways; timing/kinetics of signal transduction), caspase-3 activation, cell surface proteins and receptor internalization, RNAi library screens, cell proliferation, etc.

On-Cell Western Blot (On-Cell ELISA™) 

  • Fixed but unpermeabilized cells are stained with antibodies against extracellular protein domains, so only cell surface antigens are detected. Cell surface receptors are detected with Odyssey Infrared Imaging System. 
  • Our assay can quantify target proteins localized to the cell surface, enabling quantitative monitoring of cell surface protein expression.
  • This assay can be a high throughput assay without using radioactive labeling, to quantify ligand binding to cell surface receptors, monitor receptor internalization, and evaluate of drug effects on protein trafficking, etc.

In-Gel Western Blot  

  • Detect proteins directly in the polyacrylamide gel, without membrane transfer or blocking.
  • Similar to our Simple Western assay, In-Gel Western is useful for: large, hydrophobic, or post-translationally-modified proteins, such as glycoproteins, receptors, or cell membrane proteins that do not transfer well, and small proteins that may pass through the membrane during transfer
  • This assay also saves time, reduces cost, and eliminates variables associated with membrane transfer and blocking

      Digi-Western Array 

      • A high-throughput antibody-based microarray combining SDS-PAGE based protein separation with immobilization on Luminex beads.
      • The combination of MW resolution, sensitivity and signal linearity on Luminex platform enables the rapid quantification of hundreds of specific proteins and protein modifications in complex samples.
      • Parallel profiling of currently up to 600 total and phosph proteins in just 50 ug of samples (cells, tissues, …) to support biomarker discovery pathway mapping, drug MoA studies, lead compound characterization, etc.