( of multiple dedicated low pulsation ...)New Tools for Making and Processing Protein Microarrays.

of multiple, dedicated, low pulsation, peristaltic pumps. This enables selective blending between three bulk liquid channels, allowing use of stock buffer concentrates that can be mixed to user-specified concentrations. This removes the need to make up different buffers in advance when developing assays, allows for rapid method development, and supports the facility to perform gradient washes over the protein microarray surface, thus allowing unique flexibility in assay design. The pumps can be programmed with user definable flow rates (50 μL to 400 μL per minute, equivalent to 1 to 8 volume changes per minute across the biochip) enabling assays incorporating high affinity antibodies to be processed with higher flow rates and shortened assay times (Figure 2).

The TSA Module on the ProteinArray Workstation enables automation of a series of novel protocols that PerkinElmer Life Sciences has developed to boost the sensitivity of protein array-based assays1,2. These multi-step protocols provide for signal amplification through the use of tyramide derivatives as a substrate for horse radish peroxidase (HRP). This catalyzes a reaction to form activated tyramides which rapidly bind to multiple tyrosine residues in any proteins that are immediately adjacent to the HRP conjugate. Using a series of different tyramide derivatives and antibody detection systems, amplification can be achieved with virtually any protein assay. The amplification provides for an increase in sensitivity of up to 100-fold providing researchers with the ability to detect low abundance proteins in complex mixtures, including clinical samples.

When performed manually, the multi-step TSA assays can be very time consuming and laborious. To automate this process the ProteinArray Workstation includes the TSA Module. This automates the delivery of up to 6 reagents, including the TSA chemistries, to each of the protein arrays being processed. The module can also be used
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