ProtoArray® Human Protein Microarrays can be easily read with most commercially available fluorescent microarray scanners. See Table 1 for a list of compatible scanners.

Detection method for fluorescent applications

The high-sensitivity, low-background, signal stability, and commercial availability of fluorescence microarray scanners make fluorescence detection the preferred method for detecting interactions on the microarray. Our Molecular Probes® Alexa Fluor® detection system is the recommended fluorescent detection method. The Alexa Fluor® 647 fluorophore is brighter and more stable than other commercially available dyes such as Cy®5 dyes and is more sensitive for detecting interactions on protein arrays. We have demonstrated that detection with Alexa Fluor® 647 produces approximately 2‑fold higher signal/background ratios than Cy®5 detection. A list of known, compatible scanners is shown in Table 1. If you have any questions about the compatibility of your scanner with the ProtoArray® Human Protein Microarrays please contact technical support.

Table 1. Scanner Compatibility With ProtoArray® Human Protein Microarrays

Compatible Not Compatible
LiCor Odyssey® CLx Affymetrix GeneChip® Scanner 3000
LiCor Odyssey® SA Agilent DNA Microarray Scanner
Molecular Devices GenePix® 4000A Alpha Innotech AlphaArray®*
Molecular Devices GenePix® 4000B AppliedPrecision® arrayWoRx® 4-color Biochip Reader*
Molecular Devices GenePix® Professional 4200A Telechem SpotLight*
Molecular Devices GenePix® 4300A  
Molecular Devices GenePix® 4400A  
Perkin Elmer ScanArray® Express*  
Perkin Elmer ScanArray® Express HT*  
Perkin Elmer ScanArray® Lite  
Tecan LS Series Laser Scanner  
Tecan PowerScanner  
*Compatibility or non-compatibility is based on comparison with other like scanners

Detection method for radiolabeled applications

Detection is performed with a phosphorimager or conventional autoradiography film. The phosphorimager or autoradiography film must have resolution of at least 50 micron. The silver grain on most X-ray film is smaller than 50 micron and most common desktop scanners allow for at least 50 micron resolution scanning. For tritiated small molecule profiling, a tritium-sensitive phosphorscreen should be used.

The typical exposure time for kinase substrate profiling is 16–18 hours. For tritiated small molecule and methlytransferase substrate profiling assays the typical exposure time is two weeks. A minimum resolution of 600 dpi should be used when acquiring data with a phosphorimager.