E-Series: Competition-Based Antibody Arrays

Broad-Species Competition-based Antibody Array for Quantification or Screening

Recent technological advancements at RayBiotech have broadened the options for multiplex protein detection even further.  The RayBio® Competition-based Antibody Array (“E-Series”) is a novel array based on the competitive immunoassay principle used in our EIA kits.  In this platform, a cocktail of biotin-labeled peptides is mixed with the sample and incubated with the array; the peptides and target proteins then competitively bind to an array of capture antibodies on a solid support. Like the sandwich-based arrays (C-Series, G-Series, and Quantibody), the competition-based array can be either quantitative or semi-quantitative. If quantitative data is desired, a cocktail of peptides (derived from the target protein sequences) serves as the standard mix for derivation of calibration curves.

The E-Series array has several important advantages.  First, it enables the detection of proteins or small peptides for which no antibody pair exists, or for which antibody pairs cannot be developed.  Like the EIA kits, this array requires only 1 antibody per target for signal output.  Thus, the E-Series can detect targets not included in our sandwich-based array antibody pool.

Second, due to the high sequence homology of the peptide antigens, the E-Series antibodies have broad-species reactivity. The E-Series arrays are validated for rat, mouse, and human samples and have expected reactivity with many other mammalian species.

Third, this technology has enabled the very first quantitative antibody array that is available on both membrane and glass slide.

This novel array technology is patent-pending and available exclusively at RayBiotech.

Features

  • Picogram to nanogram LOD’s
  • High specificity and reproducibility
  • Especially useful for detection of low molecular weight targets
  • Choice of detection systems: chemiluminescence or fluorescence
  • Diversity of species detected (human, mouse and rat)
  • Diverse sample compatibility (serum, plasma, culture medium)
  • Rapid, simple procedure; only 6 hours to complete experiment
  • Low sample volume required

How It Works

E-Series: How it Works

10 different anti-adipokine antibodies are spotted onto membranes in an array format. A cocktail of 10 biotin-labeled adipokine peptides is spiked into the samples, and the labeled and unlabeled targets competitively bind to their corresponding antibodies.  After a wash step, streptavidin-horseradish peroxidase (SA-HRP) is incubated with the arrays, which interacts only with the biotin-labeled adipokines bound to the antibody.  The chemiluminescent array signals are directly proportional to the amount of biotinylated peptide-SA-HRP complex and inversely proportional to the amount of adipokines in the samples.

Research Applications

  • High-throughput profiling of adipokine & peptide hormone expression
  • Finding biosignatures for obesity, metabolism, inflammation and cancer
  • Monitoring biomarker levels in clinical trials• Identifying potential molecular targets for drug development
  • Identifying the molecular mechanisms of drug action
  • Discovering biomarkers for disease management
  • Discovering expression patterns for molecular classification of diseases

Contents of Kits

Semi-Quantitative E-Series: Membrane

EAA-ADI-E1

  • Antibody Array membranes (2 or 6)
  • 1X Blocking Buffer
  • 20X Wash Buffers 1 and 2
  • Biotinylated Peptide Cocktail
  • 500X HRP-Streptavidin
  • Detection Buffers C and D
  • 8-well washing tray
  • Plastic sheets
  • User manual

Quantitative E-Series: Membrane

QEA-ADI-E1

  • Antibody Array membranes (4 or 8)
  • 1X Blocking Buffer
  • 20X Wash Buffers 1 and 2
  • Biotinylated Peptide Cocktail
  • Standard Peptide Cocktail
  • 500X HRP-Streptavidin
  • Detection Buffers C and D
  • 2X Lysis buffer
  • Protease inhibitor cocktail
  • 8-well washing tray
  • Plastic sheets
  • User manual