C-Series: Membrane-Based Antibody Arrays

Semi-Quantitative, Sandwich-Based, Membrane Antibody Arrays

C-Series Membrane-based Antibody Array

The RayBio® Membrane-Based 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.

The C-Series 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.

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 Antibody 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 array kits are available in-stock, detecting anywhere from 10 to 274 proteins simultaneously.

For a comparison & overview of all our antibody array systems, click here.

Compare C-Series Protein ListsCustomize Your ArrayBrowse C-Series 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

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

References

  1. Ray S, Britschgi M, Herbert C, Takeda-Uchimura Y, et al. Classification and prediction of clinical Alzheimer's diagnosis based on plasma signaling proteins. Nature Med. 2007; 13(11):1359-1362.
    Species: Human
    Sample Type: Plasma
  2. Marks DJB, Harbord MWN, MacAllister R, Rahman FZ, Young J, et al. Defective acute inflammation in Crohn's disease: a clinical investigation. Lancet. 2006;367:668-678.
    Species: Human
    Sample Type: Other Fluid
  3. Wang D, Wang H, Brown J, Daikoku T, Ning W,et al.CXCL1 induced by prostaglandin E2 promotes angiogenesis in colorectal cancer. J Exp Med. 2006; 203:941-951
    Species: Human
    Sample Type: Conditioned Media
  4. Chou C, Chen P, Lee P, Cheng A, Hsu H, et al. Radiation-Induced Hepatitis BVirus Reactivation in LiverMediated by the Bystander Effect from Irradiated Endothelial Cells. Clin Cancer Res. 2007;13;851–857.
    Species: Human
  5. Sohara Y, Shimada H, Minkin C, Erdreich-Epstein A, Nolta J,et al. Bone Marrow Mesenchymal Stem Cells Provide an Alternate Pathway of Osteoclast Activation and Bone Destruction by Cancer Cells. Cancer Res. 2005;65(4):1129-1135
    Species: Human
    Sample Type: Conditioned Media
  6. McAllister SS, Gifford AM, Greiner AL, Kelleher SP, Saelzer MP, et al. Systemic Endocrine Instigation of Indolent Tumor Growth Requires Osteopontin. Cell. 2008;133:994-1005.
    Species: Human
    Sample Type: Plasma
  7. Zhang J., et al. YAP-dependent induction of amphiregulin identifies a non-cell-autonomous component of the Hippo pathway. Nat Cell Biol. 2009 Dec;11(12):1444-50. doi: 10.1038/ncb1993.
    Species: Human
    Sample Type: Cell Lysate
  8. Kang TW., et al. Senescence surveillance of pre-malignant hepatocytes limits liver cancer development. Nature. 2011 Nov 9;479(7374):547-51. doi: 10.1038/nature10599.
    Species: Mouse
    Sample Type: Tissue Lysate
  9. Carnevalli L., Scognamiglio R., Cabezas-Wallschied N., Rahmig S., Laurenti E. Improved HSC reconstitution and protection from inflammatory stress and chemotherapy in mice lacking granzyme B. Journal of Experimental Medicine vol. 211 no. 5 769-779
    Species: Mouse
    Sample Type: Conditioned Media
  10. Stanford J. Efferocytosis produces a prometastatic landscape during postpartum mammary gland involution. JCI Sept 2014, epub ahead of print. doi:10.1172/JCI76375.
    Species: Mouse
    Sample Type: Tissue Lysate
  11. Braig M., Pallman N., Preukschas M., Steinemann D., Hofmann W., et al. A ?telomere-associated secretory phenotype? cooperates with BCR-ABL to drive malignant proliferation of leukemic cells. 2014 Leukemia, Pubslihed April 1, Epub Ahead of Print, doi: 10.1038/leu.2014.95
    Species: Mouse
    Sample Type: Conditioned Media
  12. Deuse T., Hua X., Wang D., Maegdefessel L., et al. Dichloroacetate prevents restenosis in preclinical animal models of vessel injury. Nature (2014) doi:10.1038/nature13232 [Epub ahead of print]
    Species: Rat
    Sample Type: Tissue Lysate

More references are listed per product on the individual product pages.