ProPlate® Chambers for Macroarray Printing
Many laboratories desire the benefits of utilizing microarray technology in their field of interest but do not have access to microarray printing equipment or cannot justify the expense of dedicated print runs for lower throughput experiments. In many cases where highly multiplexed assays are not a necessity or during early-phase discovery phases of research projects microarray printing is not required and utilization of “macroarrays” is often sufficient to address the biological question. ProPlate® Chambers offer a simple and robust method for depositing macroarrays in a consistent manner onto binding substrates without the aid of automated or manual pin-based spotting devices. As a demonstration of the effectiveness of this system, shown below is a macroarray spotted onto a nitrocellulose film slide utilizing a 64-well ProPlate® Chamber and used for an assay optimization application.
Figure 1. Macroarrays utilized for timecourse and dynamic range experiments
Figure 1. Shown are 5 macroarrays spotted in triplicate per nitrocellulose film slide using 64-pad ProPlate® Chambers. The spotting layout is outlined in the legend. Column A contains a dilution of goat serum spiked with anti-goat IgG-TRITC in decreasing concentration from Rows 1- 5. Columns B-D were spotted with un-labelled goat serum (0.1, 1, 10 mg/ml, respectively) and assayed with anti-goat IgG at various titers (rows 1-5: 100, 10, 1, 0.1, 0.01 µg/ml). Arrays were used to determine optimal protein binding times (protein solutions allowed to sit in the ProPlate® wells for increasing periods of time) as well as optimal assay concentrations and optimal assay incubation times for a simple binding assay (also performed in the ProPlate® wells).
Figure 2. Results from Experiments Performed in ProPlate® Chambers.
Figure 2. Shown are some of the data obtained with this experiment. The macroarray image in Panel A is a compilation of all data obtained from Column A from all arrays (Fig. 1) looking at protein binding dynamics on an AVID nitrocellulose film slide. The data corresponding to this image is shown in Panel B showing linearity of protein binding to the AVID surface to approximately 5 mg/ml protein concentration. Panel C shows results from the functional assay performed with the spotted macroarrays. Reported is the dynamic range of the assay when using various primary antibody titers (100, 10, 1, 0.1, 0.01 µg/ml) with spotted material deposited at 3 spotting concentrations (0.1, 1, or 10 mg/ml).