Proteins are the most abundant and integral molecules in all organisms, playing an important role in almost every cellular process. As such, they must be studied and analyzed in order to understand them better. In recent years, advancements in proteomic tools have allowed scientists to gain insight into a wide array of protein-related information with greater efficiency. One tool that has become increasingly popular for protein profiling is antibody arrays such as cytokine arrays, to measures many cytokines at the same time using a single sample. These innovative technologies offer scientists the ability to measure changes in a range of proteins all at once without sacrificing accuracy or quality results. In this blog post, we’ll explore these proteomic tools further and discuss their various applications and uses so you can leverage them for your next research project!

Overview of cytokine arrays and their importance in research

Proteomic tools such as cytokine arrays offer a powerful set of strategies for understanding complex biological systems, providing researchers and developers with unparalleled insights into the functional properties of proteins.

What is a cytokine array?

Cytokine arrays offer a powerful tool for molecular analysis that can be used to further our understanding of complex biological systems. Cytokine arrays enable multiplex protein profiling from multiple species options such as human, mouse, rat, rabbit, porcine, and bovine cytokine arrays. This allows researchers to rapidly identify and measure cytokines – proteins responsible for communication between cells – from a single sample e.g., serum, plasma, cell culture supernatant, lysates, or other biological fluids.

Advantages of multiplex protein profiling using cytokine arrays

Cytokine arrays are a multiplex assay that measures the expression of multiple proteins at the same time. They allow a much broader view of protein activity than can be obtained with single-target ELISA assays and western blots. These arrays allow for a more comprehensive analysis of over 6,000 proteins, encompassing a wide variety of cytokines, chemokines, growth factors, receptors, adipokines, proteases, signaling and structural proteins, and metabolic enzymes. Obtain a broad view of relevant biological processes like inflammation, apoptosis, angiogenesis, as well as identification of key factors of disease mechanisms in cancer, infectious disease, cardiovascular disease, and more for efficient protein biomarker research. Depending on the sample or analyte requirements, several techniques for visualizing protein expression (chemiluminescent, fluorescent, flow cytometer, etc.) can be used for a comprehensive analysis of biological responses and functions and identification of key factors or disease mechanisms.

 

Overview of the types of cytokine arrays

Cytokine arrays provide a powerful tool for investigating complex biological pathways in biomedical research. Depending on the experiment, researchers have the choice of membrane-based, glass-based, or label-based cytokine arrays. Membrane-based cytokine arrays offer a broad detection range with minimal sample and labeling requirements, while glass-based setups are ideal for high resolution purposes since the flat surface allows for tight control on spot size and distance. Label-based cytokine arrays provide convenient convenience testing and analysis capabilities. As such, each type of array offers unique advantages that allow scientists to choose the most suitable approach suited to their particular use case.

Membrane-based cytokine arrays

Membrane-based cytokine arrays use a panel of capture antibodies printed on a nitrocellulose membrane. The protein from the incubated sample is captured, and a detection antibody is also bound, creating an antibody-antigen-antibody sandwich. Chemiluminescent signals are then visualized on x-ray film or a digital image, allowing calculation of the relative amount of protein present. C-series arrays are processed similarly to a western blot (chemiluminescent readout) and are simple enough for a researcher to successfully collect robust data with little or no optimization.

Glass slide-based cytokine arrays

Glass slide-based cytokine arrays also utilize the sandwich immunoassay principle but require much less sample than membrane-based arrays. The capture antibodies are spotted on 75mm x 25mm glass slide solid supports and a corresponding biotinylated detection antibody is utilized for resolving fluorescent signals. Each slide is spotted with either 8 or 16 identical antibody arrays (also called “subarrays”). They are available as Quantitative / Multiplex ELISA (called Quantibody® arrays), which are quantitative, glass slide multiplex ELISA cytokine arrays with cytokine standards. They are also available as a semi-quantitative, glass-based arrays (called G-Series Arrays) that measure relative protein expression levels across samples or experimental conditions. Unlike Quantibody®, G-series arrays do not come with cytokine standards.

Label-based cytokine arrays / High throughput cytokine arrays

Label-based cytokine arrays differ from sandwich ELISA-based arrays primarily in the means of detection. Instead of using a second biotinylated antigen-specific antibody, sample proteins themselves are labeled with biotin prior to incubation with the capture antibodies. This prevents unintended antibody interactions thus eliminating limitations on the size of the array panel. Available on both glass slide and membrane formats, L-Series arrays are ideal for biomarker discovery studies and exploratory, high-throughput screening.

What is a Human Cytokine Array & How Can It Help You With Protein Profiling

A human cytokine array is a research array platform that is used to measure the expression of cytokines in human cells or animal cells. Cytokines are proteins that play a role in cell signaling, and they can be used to determine the health of a patient’s immune system. Human cytokine arrays offer many advantages over other methods of protein profiling, including greater precision and the ability to detect low-level signals. Here, we will discuss the benefits of human cytokine arrays for protein expression profiling.

A human cytokine array is a powerful tool for protein profiling. It involves immobilizing a panel of different human cytokines onto a solid surface and then monitoring the interactions that occur between those proteins and any given sample. In this way, one can identify a comprehensive profile of the proteins present in the sample, determine their relative concentrations as well as measure biochemical interactions. A great advantage to using human cytokine arrays is the ease with which researchers can access large amounts of data with just one single measurement. As seen in many research publications, they are extremely useful for studying host-pathogen interactions, immune regulation, and other important biological processes. There are many useful tools of RayBiotech’s human cytokine arrays for the study of human cytokines, as well as chemokines, growth factors, angiogenesis, apoptosis, and other proteins using the different types of cytokine arrays.

Uncover new insights with Mouse Cytokine Arrays

The mouse has long been a model organism in biomedical research, and a tool for studying the effects of drugs and disease. In particular, mouse cytokine arrays have become indispensable in studying the immune system. We explore the potential of mouse cytokine arrays in research and how they are helping to shape our understanding of human health and disease.

RayBiotech offers a variety of mouse cytokine arrays that allow researchers to study the immune response and other biological processes in great detail. These arrays are designed to detect changes in mouse cytokine levels, making them ideal for studying various diseases and conditions. Cytokine arrays are available for multiple species options: Human, Mouse, Rat, Rabbit, Porcine, and Bovine cytokine arrays. Browse our full offering by using our species filter on the following platform product searches.

C-Series Membrane Arrays: Semi-Quantitative, Sandwich-based, Membrane Antibody Arrays

RayBiotech’s 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 (usually 2.5 cm x 3 cm). The array membranes are processed similarly to a Western blot (chemiluminescent readout).

Quantibody: Quantitative, Sandwich-based, Glass Slide Antibody Arrays

Quantibody® is an array-based multiplex ELISA system for simultaneous quantitative measurement of multiple cytokines, growth factors, proteases, soluble receptors, and other proteins in a wide variety of sample types. Quantibody combines the high specificity and sensitivity of ELISA with the high throughput of the glass chip-based array. With this platform, only 50 µl of sample is needed for quantification of up to 40 proteins in quadruplicate, making this array 80 times more efficient than traditional ELISA.

 

Applications of Cytokine Arrays in Research

Cytokine arrays have become increasingly popular for research use, with their applications spreading from cancer research to neuroscience studies. Researchers often rely on cytokine arrays to test cytokines in tissue samples, blood, and other fluids, making cytokine analysis more straightforward than ever before. The results of cytokine array analyses are regularly published in scientific publications, which further highlights the utility of this approach. Browse our interactive search table of publications to learn how cytokine array kits are used to advance biomarker discovery research. Get answers to your questions, such as:

• How often has this kit been cited?
• Has my sample type been used with this kit?
• How have other researchers prepared my sample type for this assay?

Browse all citations using RayBiotech’s cytokine arrays to advance research

• Learn how RayBiotech cytokine arrays are used in research

Examples of research publications that have cited cytokine arrays for these applications

 

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 signalling 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.

 

Future Directions for Protein Profiling with Proteomic Tools

Proteomic research tools continue to be at the forefront of research in molecular biology. While much research has already been conducted using cytokine arrays for protein profiling applications, numerous research publications indicate there is much more research needed to be done and offer many innovative ways research can take advantage of proteomic techniques for protein profiling purposes. Researchers strive to uncover new pathways and mechanisms of action through which these techniques may be further utilized and expand on previous research citations that have pointed out the potential for cytokine arrays in protein profiling studies. Developing an understanding of how proteomics affects diverse physiological properties and results will aid progress for countless areas within the field of molecular biology. For enquiries on cytokine arrays for research, contact info@genomax.com.sg if you would like to learn more about how Genomax Technologies can help make your project a success.