Using Antibody Arrays for Fast Cytokine Profiling

The amount of multiple different cytokines in a sample can be measured simultaneously and easily using a cytokine targeting antibody array.

Abcam’s membrane cytokine arrays, like their other antibody arrays, allow up to 96 cytokines, or similar proteins, in a sample to be determined from one experiment.

The arrays have a sensitivity of a pictogram per millimeter making them ideal tools to investigate proteins expressed at low levels. Like an ELISA, the assays are based on antibodies and use a similar procedure and equipment as a chemiluminescent western blot.

In addition to cytokine analysis, the assays can also be used to analyze the levels of other proteins such as growth factors, chemokines, and proteins associated with angiogenesis and apoptosis and amongst others. Examples of Abcam’s arrays are cytokine array ab133997 and cytokine array ab133998, which can be used for the measurement of up to 42 and 80 human cytokines respectively.

The Benefits of Using Membrane-Based Arrays for Other Targets

Antibody arrays reduce both the time and economic cost required for the simultaneous measurement of multi-protein samples. Using arrays prevents researchers from having to run individual western blot or ELISA experiments for each analyte.

Antibody arrays provide the following benefits:

  • The detection method (chemoluminescence) is the same as western blot methods
  • The arrays can be used with existing western blot recording systems
  • No specialist equipment is needed
  • Free analysis software for densitometry and spot recognition is provided – available for download online
  • A large range of detection (10,000-fold in picograms per milliliter)
  • The kit is optimized, containing detection antibodies, membranes, and buffers
  • Duplicate antibody spots mean captured antibodies are spotted in duplicate

How are Membrane-Based Arrays Carried Out?

Antibody arrays function in a similar way to ELISA, another antibody-pair based assay, though antibody arrays use a membrane substrate and not a plate. Antibodies are spotted over the membrane to generate the array, with every pair of spots acting as a sensor for a particular analyte.

To carry out an analysis, 0.2 – 1 mL of sample is added to the membrane, followed by streptavidin-HRP and biotinylated detector antibodies. The entire assay can be easily run overnight or in a day.

Analysis of the array then takes similar steps to the western blot method. Comparing samples is either achieved by densitometry software, for a semi-quantitative result or by eye.

Comparing the Different Techniques

ELISA Membrane antibody arrays
No of proteins measured 1 up to 96 per array
Cost per sample Low ~1/2 cost of ELISA on a per sample per analyte basis
Sensitivity pg/ml pg/ml
Quantitative Yes Semi
Sample volume ~0.1 mL 0.2-1 mL
Protocol length/ complexity Similar Similar
Required equipment Plate reader Same as for any chemiluminescent western blot

Typical Applications of Cytokine Arrays

  • Profiling biomarkers present in serum samples
  • Profiling the different cytokines present during the inflammatory response
  • Profiling cytokines in normal, diseased, and treated samples
  • Confirming the results of western blotting, gene microarrays, or PCR experiments
  • Determination of biological events, e.g., those in asthma

About Abcam

Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.

Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.

Sponsored Content Policy: publishes articles and related content that may be derived from sources where we have existing commercial relationships, provided such content adds value to the core editorial ethos of News-Medical.Net which is to educate and inform site visitors interested in medical research, science, medical devices and treatments.

Last updated: Apr 1, 2019 at 6:23 AM


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