Antibody Arrays

More Data, Less Sample

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. 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.

C-Series

C-Series Array

Detection Chemiluminescent
Solid Support Membrane
Design Principle Sandwich ELISA
Results Semi-quantitative

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Quantibody®

Quantibody Array

Detection Fluorescent
Solid Support Glass Slide
Design Principle Sandwich ELISA
Results Quantitative

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G-Series

G-Series Array

Detection Fluorescent
Solid Support Glass Slide
Design Principle Sandwich ELISA
Results Semi-quantitative

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E-Series

E-Series Array

Detection Chemiluminescent
Solid Support Membrane
Design Principle Competitive ELISA
Results Semi-quantitative or Quantitative

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L-Series

L-Series Arrays

Detection Fluorescent or Chemiluminescent
Solid Support Glass Slide or Membrane
Design Principle Direct Labeling (Biotin)
Results Semi-quantitative

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Isotyping

Isotyping Arrays

Detection Fluorescent or lateral flow
Solid Support Glass Slide
Design Principle Sandwich ELISA
Results Qualitative

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Comparison of Antibody Array Platforms

NAME APPLICATIONS SOLID SUPPORT DESIGN PRINCIPLE OUTPUT # OF ANALYTES SPECIES*
Quantibody® protein expression profiling slide sandwich-based quantitative 10 – 440 H, M, R, B, C, F, E, P, L, N
C-Series Arrays protein expression profiling membrane sandwich-based semi-quantitative 10 - 274 H, M, R
G-Series Arrays protein expression profiling slide sandwich-based semi-quantitative 10 - 274 H, M, R
L-Series Arrays protein expression profiling both label-based semi-quantitative 90 - 1000 H, M, R
Phospho Arrays phosphorylation profiling both sandwich-based semi-quantitative 17 - 71 H
E-Series Arrays protein expression profiling membrane competition-based quantitative 10 H
Lectin Arrays protein-lectin interaction.
Meaning?... Which protein-bound sugars are present in my sample?” or “which sugars are bound to my protein of interest?
slide label-based semi-quantitative 40 any
Glycome Arrays glycosylation profiling.
Meaning?... Which proteins in my sample are glycosylated, and to what extent?
slide sandwich-based (lectin-ab pair) semi-quantitative 507 any
Protein Arrays auto-antibody profiling, characterizing antibody specificity, More... Can be adapted for detection of protein-protein interactions, protein modifications, small molecule-protein interactions slide label-based semi-quantitative 48 - 487 H, M
Rapid Isotyping Strips isotyping; hybridoma screening and selection lateral flow strip sandwich-based qualitative 10 M
Rapid Isotyping Arrays isotyping; hybridoma screening and selection slide sandwich-based semi-quantitative 8 - 10 M, R

*H=human  |  M=mouse  |  R=rat  |  P=porcine  |  C=canine  |  F=feline  |  B=bovine  |  E=equine  |  N=rhesus monkey  |  L=rabbit

How To Choose An Array

YOUR SPECIFIC NEED OUR SOLUTION
I want to screen as many factors as possible (I need a "big net") L-Series: Label-based Arrays or larger Quantibody® Arrays
I want to focus on a specific pathway or biological process Pathway-specific arrays (e.g. Inflammation; Apoptosis, etc) or Phosphorylation Arrays
I want to choose a specific panel of markers Custom Array
I have a limited sample volume Glass Slide-based Arrays:
I don't have a laser scanner Membrane-based Arrays:

Or use our free glass slide scanning service

I want quantitative results Quantibody® Arrays or E-Series Arrays
I want to identify antibody isotypes Isotyping Arrays
I want to study protein glycosylation Glycobiology Arrays (Lectin ArraysGlycosylation ArraysGlycan Arrays)
I want to screen protein-protein interactions Protein Arrays
I have samples from an uncommon species L-Series: Label-based Arrays or Quantibody® Arrays
I want to do biomarker discovery Any RayBiotech Array
  C-Series G-Series L-Series Quantibody®
Chemiluminescence    
Fluorescence  
Equipment Needed CCD, X-ray, gel doc Laser scanner CCD, X-ray, gel doc
Laser scanner
Laser scanner
Sensitivity pg to ng pg to ng pg to ng pg to ng
Target Density Low to High Low to High Low to High Low to High
Semi-Quantitative  
Quantitative      
Specificity Very High Very High High Very High

How Arrays Can Advance Your Research

Due to the sheer number of secreted factors that could potentially be involved in cell–cell interactions, high-content screens are essential to identifying key secreted factors.

Straussman R, Morikawa T, Shee K, Barzily-Rokni M, Qian ZR , Du J, et al. Tumour micro-environment elicits innate resistance to RAF inhibitors through HGF secretion. Nature. 2012; 487: 500-504.

The investigators incubated melanoma (skin cancer cells) in vitro with cell-cultured media obtained from multiple stromal cell lines and identified those media that induced the highest degree of stromal-mediated resistance to a RAF inhibitor (recently approved to treat melanoma). They then used both the G-Series 4000 and L-Series 507 Arrays to characterize the expression profiles of secreted factors in stromal cell-cultured media that induced the greatest degree of drug resistance in the cancer cells. They then used statistical analysis of the individual secreted factors detected in the various expression profiles to find those that correlated best with drug resistance.

High-density screening arrays help you determine where to focus to get the biggest return on your research efforts.

Scheel C, Eaton EN, Li SH, Chaffer CL, Reinhardt F, Kah K-J, 2, Bell G, Guo W, Rubin J, Richardson AL, Weinberg RA. Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast. Cell. 2011; 145(6):926-940.

This paper focuses on the mechanism of epithelial-to-mesenchymal transition (EMT), which is an important process in tumor formation. The Human L-Series 507 Array yielded a short list of targets, including the TGFβ and WNT/β-catenin pathways, which are known to antagonize one another. By knowing where to look, they were able to focus their efforts on the right signal pathways, greatly improving their chances of working out the entire mechanism of this process, which netted them a paper in Cell.

Using a cytokine array keeps you from missing something important.

Vargas DL, Nascimbene C, Krishnan C, Zimmerman AW, Pardo CA. Neuroglial activation and neuroinflammation in the brain of patients with autism. Annals of Neurology. 2005; 57(1):67–81.

The authors of the paper found the first evidence of a neuroinflammatory process in the pathology of autism, even identifying several key factors that pointed to the mechanism of this process. This neuroinflammatory component of autism had been suspected for years, but no one had ever proven it. If the authors had decided to select a few ELISA kits or even a 20-plex bead panel to do this investigation instead of an array kit detecting 120 different cytokines, the authors may have missed identifying the key targets, and we might still be wondering if autism has a neuroinflammatory component.

Cell–cell signaling pathways are complex, and multiple factors may be working together, in concert. Therefore, screening for a limited number of secreted targets will often give you an incomplete picture.

Coppé J-P, Patil CK, Rodier F, Sun Y, Muñoz DP, Goldstein J, Nelson PS, Desprez P-Y, Campisi J. Senescence-associated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biology. 2008; 6(12).

This paper describes the first characterization of the senescence-associated secretory phenotype in stromal–tumor interactions. It uncovers the expression profile of permanently senescent tissues (epithelial and stromal) that secrete a wide range of factors that included pro-inflammatory factors, growth factors, cell-adhesion factors and proteases that restructure the extracellular matrix (ECM). In subsequent inquiries, these various components of this broad secretory profile were shown to work in concert to promote tumorigenesis. The Human Cytokine Array 1000 was instrumental in this discovery.