| Solution Information | help | |
| Enzyme: | Mitogen-activated protein kinase kinase kinase kinase 2 | |
| inhibitor: | BDBM51801 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Similar protocol was utilized as in the primary screening campaign. For each target, the components of the multiplex consist of a Streptavidin-functionalized polystyrene bead (Spherotech, USA), a biotinylated MEKK-fusion protein target (3 total, supplied by assay provider), and a fluorescent peptide probe (AlexaFluo488-GST-MEK5 WT, supplied by the assay provider). Beads for each target have been labeled with varying intensities of red color, such that each assay is built on a unique bead set, and each bead set is associated with a unique optical address. The similarly sized bead sets (~5 micron) are distinguished by distance emission characteristics at 750 nanometer with excitation at 635 nanometer. Bead sets are coated with biotin-conjugated MEK kinase protein targets (final protein concentration 1 microM) in HPSN bead coupling buffer (30milliM HEPES, 100milliM KCl, 20milliM NaCl, 0,01% NP-40, pH 7.5) at 4 degrees C, and incubated overnight (16-18 hours). The three bead sets (each | |
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