Membrane Antibody Arrays: C-Series

Semi-Quantitative, Sandwich-Based, Membrane Antibody Arrays

C-Series Membrane-based Antibody Array

The RayBiotech Membrane Antibody Arrays (C-Series™) are tools for screening and comparing expression levels of many cytokines, growth factors, proteases, soluble receptors, and other proteins in a wide variety of sample types.

Many biological processes such as apoptosis, inflammation, angiogenesis, immune response and migration often accompany changes of cytokine expression levels. Because of the extensive cross-talk between cytokines, a complete analysis of biological responses and functions must be obtained through multiplex assays. The C-Series membrane arrays allow a much broader view of protein activity than can be obtained with single-target ELISAs and Western blots. Moreover, antibody array screening improves the chances for discovering key factors, disease mechanisms or biomarkers related to cytokine signaling.

A variety of C-Series membrane array kits are available in-stock, detecting anywhere from 10 to 274 proteins simultaneously. For higher-density screening membrane arrays, check out our L-Series: label-based antibody arrays.

Features

  • Simultaneous detection of multiple proteins
  • High specificity (sandwich antibody pair used)
  • High sensitivity (pg/ml levels)
  • No dedicated equipment required; compatible with any chemiluminescent imaging system
  • Easy to use, no training required
  • Results can be obtained in 1 day

Contents of Kit

  • RayBio® Antibody Array Membranes
  • Biotinylated Detection Antibody Cocktail
  • Blocking Buffer
  • Wash Buffers 1 and 2
  • Cell & Tissue Lysis Buffer
  • Detection Buffers C and D
  • Plastic Incubation Tray
  • Protease Inhibitor Cocktail (in select kits)
  • Manual

How It Works

C-Series Antibody Arrays: How it works
The C-Series membrane arrays utilize the sandwich immunoassay principle, wherein a panel of capture antibodies is printed on a nitrocellulose membrane solid support (usualy 2.5 cm x 3 cm). The array membranes are processed similarly to a Western blot (chemiluminescent readout). Signals are then visualized on x-ray film or a digital image, allowing densitometry data collection and calculation of fold-changes for each detected protein. The entire procedure can be completed in 1 day, and is simple enough to permit even the novice researcher to successfully collect data with very few pitfalls and little or no optimization.

Handling The Membranes

Research Applications

  • High-throughput profiling of cytokine expression
  • Identifying potential molecular targets for drug development
  • Identifying the molecular mechanisms of drug action
  • Identifying crucial factors involved in disease processes
  • Discovering biomarkers for disease management
  • Discovering expression patterns for molecular classification of diseases

Technical Notes

Comparison to ELISA

  • More Data, Less Sample: Antibody arrays provide high-content screening using about the same sample volume as for ELISA.
  • Global View of Cytokine Expression: Antibody array screening improves the chances for discovering key factors, disease mechanisms or biomarkers related to cytokine signaling.
  • Greater Sensitivity: As little as 4 pg/ml of MCP-1 can be detected using the C-Series array format. In contrast, our similar MCP-1 ELISA assay has a sensitivity of 40 pg/ml of MCP-1.
  • Increased Range of Detection: ELISA assays typically detect a concentration range of 100- to 1000-fold, however, RayBiotech arrays can detect IL-2 at concentrations of 25 to 250,000 pg/ml, a range of 10,000-fold.
  • Better Precision: As determined by densitometry, the inter-array Coefficient of Variation (CV) of spot signal intensities is 5-10%, comparing favorably with ELISA testing (CV = 10-15%).

Features

  • High Sensitivity: Can detect as low as 1 pg/ml for some proteins.
  • Positive control spots: To orientate each membrane, monitor the detection steps, and normalize the results.
  • High Reproducibility: Inter-array Coefficient of Variation (CV) of spot signal intensities as low as 5% when run under optimal conditions.
  • High Specificity: A recombinant protein for each corresponding antibody spot is tested to ensure that only one particular spot is visible after hybridization.
  • High Quality: Many antibody pairs are tested for each detectable protein target but only the pairs which show the strongest affinity and sensitivity are selected for the array.
  • Wide Dynamic Range: A detectable range as high as 4 orders of magnitude (104) for some proteins.
  • Proven Technology: There are hundreds of publications using C-Series antibody arrays.
  • Robust Data Output: Each antibody is printed in duplicate providing multiple data points for each detected protein target.
  • Small Membrane Size: Average membrane dimensions only 2.5 X 3.0 cm
  • Duplicate Spots: The antibodies printed on the C-Series arrays are spotted in duplicate, decreasing the number of samples needed to run, saving precious sample volume while still allowing for robust data output.
  • Negative Control and Blank spots: To assess baseline and background responses.
Sample Images
IL-6 Membrane Image