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Antibody Arrays

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 Comparison

G-Series Comparison

L-Series Comparison

Quantibody Specifications

E-Series Comparison

C-Series Antibody Arrays

The C-Series arrays feature chemiluminescent signal detection. The antibodies are spotted on nitrocellulose membrane solid supports and are handled in a very similar manner to Western blots. 

All C-Series arrays work on the sandwich ELISA principle, utilizing a matched pair of antibodies: an immobilized capture antibody and a corresponding biotinylated detection antibody.

Working sample volume: 1,000 µl (1 ml)

Advantages & Features:

  • No specialized equipment required
  • Easy to use
  • High specificity
  • Low background, high sample diversity, etc.

Learn More      Browse C-Series Arrays

 

G-Series Antibody Arrays

The G-Series arrays feature fluorescent signal detection. The antibodies are spotted on glass slide solid supports and require a laser scanner for data collection.

All G-Series arrays work on the sandwich ELISA principle, utilizing a matched pair of antibodies: an immobilized capture antibody and a corresponding biotinylated detection antibody.

Working sample volume: 50-100 µl

Advantages & Features

  • Lower sample consumption
  • Easy to use; high throughput
  • Wider dynamic range (up to 5 orders of magnitude)
  • Low background, high sample diversity, etc.

Learn More      Browse G-Series Arrays

 

L-Series Antibody Arrays

The L-series arrays utilize direct labeling for signal detection, wherein the antigen is tagged with biotin prior to incubation with the capture antibody.  The signal is then developed with a streptavidin-conjugated HRP or fluor.  Since this array requires only a single antibody per target molecule (as opposed to an antibody pair), any possibility of interactions between antibodies within the same array panel is eliminated.  Thus, an unlimited number of antibodies may theoretically be included in each panel, making this array platform ideal for high-content screening of protein expression.

The capture antibodies for L-Series may be spotted on either glass slide or membrane.

Working sample volume: 20-100 µl 

Advantages & Features

  • Hundreds of molecules can be screened simultaneously
  • Lowest cost per target 
  • High specificity
  • Compatible with serum, plasma, and cell culture media

Learn More      Browse L-Series Arrays

 

Quantibody Arrays

The Quantibody arrays are quantitative multiplex ELISA arrays featuring fluorescent detection. The antibodies are spotted on glass slide solid supports and require a laser scanner for data collection. Cytokine standards are provided with the array for calculation of target protein concentrations.

All Quantibody arrays feature the sandwich immunoassay principle, utilizing an immobilized capture antibody along with a corresponding biotinylated detection antibody.

Working sample volume: 50-100 µl 

Advantages & Features

  • Lower sample consumption
  • Easy to use; high throughput
  • Wider dynamic range (up to 5 orders of magnitude)
  • Low background, high sample diversity, etc.

Learn More      Browse Quantibody Arrays

 

E-Series Antibody Arrays

E-Series arrays are the very first antibody arrays on the market to utilize the competitive immunoassay method for signal detection.

Working sample volume: inquire

Advantages & Features

  • No specialized equipment required
  • Easy to use
  • High specificity
  • Low background, high sample diversity, etc.

Learn More      Browse E-Series Arrays

 

Rapid Arrays

 

 

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?...
slide label-based semi-quantitative 40 any
Glycome Arrays glycosylation profiling. 
Meaning?...
slide sandwich-based (lectin-ab pair) semi-quantitative 507 any
Protein Arrays auto-antibody profiling, characterizing antibody specificity, 
More...
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 Arrays,Glycosylation Arrays,Glycan 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 secretionNature. 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.

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