MSD Multi-Array
Cell-based multiplex assay using MULTI-SPOT Phosphoprotein Kits are available from MSD (www.meso-scale.com). Logarithmically growing HEK293 cells were treated with or without UV and activation of JNK, p38, and p44/42 was examined using antibodies specific for the phosphorylated protein (A). Comparative, representative Western blot is shown in B for comparison. Whole cell lysates were added to MSD MULTI-SPOT 4 Spot 96-well plates coated with capture antibody at three of four spatially distinct electrodes per well. Phosphorylated proteins were detected with MSD SULFO-TAG detection antibodies.
The Meso Scale Discovery™ platform is a solid phase immunoassay methodology that uses electrochemiluminescence (ECL) for detection of bound phosphoprotein. The ECL system employs proprietary microtiter plates that include screen printed electrodes which are used as solid phase support for binding reagents and for inducing ECL. Different electrode spotting configurations available on the plates permit the arraying of several capture antibodies in a single well. Lysate of interest is added along with a cocktail of ruthenium-labeled detection antibodies. Quantification is done with either the Sector® Imager 6000 for high throughput analysis or the Sector® PR 100 Reader, an ECL reader intended for lower throughput and assay development.
MULTI-ARRAY™ is a trademark of Meso Scale Discovery.
