Membrane antibody arrays
Related
Antibody arrays such as cytokine arrays and arrays for other analytes are simple tools to measure the levels of many cytokines at the same time
Our membrane cytokine arrays and other antibody arrays enable the analysis of the levels of up to 96 cytokines or other proteins in just one experiment. These antibody arrays have pg/ml sensitivity and are easy tools for comparison of up to twenty samples in one experiment. They are antibody pair based assays, just like an ELISA, and only require the same equipment as any chemiluminescent western blot for analysis.
As well as cytokines, they are also useful tools for the study of chemokines, growth factors, angiogenesis, apoptosis and other proteins. Good examples of our cytokine arrays are cytokine array ab133997 and cytokine array ab133998, for measurement of 42 and 80 human cytokines respectively.
Advantages of membrane-based cytokine arrays and antibody arrays for other targets
Antibody arrays are used to measure many proteins simultaneously in one experiment, with significantly less time, effort and cost than running the many ELISAs or western blots required for the same number of analytes. Compared to other multiplex immunoassays, such as bead-based assays, membrane antibody arrays are very easy to set up and are significantly cheaper for comparison of when you need to compare many proteins across a small number of samples.
Antibody arrays offer several advantages:
- No special equipment required
- Chemiluminescent detection method is the same as a western blot
- Suitable for any chemiluminescent blot documentation system
- Film developer or benchtop darkroom
- Free densitometer and spot recognition analysis software
- Available for download for free from NIH
- Suitable for use with the Li-Cor® detection system Adapted array protocol for Li-Cor®
- Large detection range
- Range of 10,000 fold detection (in pg/mL)
- Duplicate antibody spots
- Capture antibodies are spotted in duplicate.
- Optimized kit
- Our antibody array kit contains buffers, detection antibodies and membranes.
How do membrane-based antibody arrays work?
Antibody arrays are an antibody-pair-based assay, analogous to ELISA, but using a membrane as a substrate rather than a plate. Capture antibodies are supplied arrayed/spotted on a membrane with each pair of spots representing a different analyte. Sample is added (0.2-1ml of 1 sample to each membrane), and then paired biotinylated detector antibodies and streptavidin-HRP.
The array is analyzed using the same methods as a chemiluminescent western blot. Comparison between samples can be by eye or using densitometry software for a semi-quantitative comparison. The whole assay can be run in a single day or with overnight incubations.
Table comparing 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 | ~0.1mL | 0.2-1mL |
| Protocol length/ complexity | Similar | Similar |
| Required equipment | Plate reader | Same as for any chemiluminescent western blot |
Example applications
- Multiplex cytokine expression profiling in inflammation
- Serum biomarker profiling
- PCR, gene microarray or western blotting results confirmation
- Detection of active biological processes, for example in inflammation, asthma
- Relative profiling of normal, diseased and treated samples