The Muse® H2A.X Activation Dual Detection kit is part of a series of products that include a pair of antibodies which bind to the same protein; one antibody detects total protein expression and the other detects the phosphorylated form of the same target.
Two-parameter analysis allows for specific detection of phosphorylation and eliminates false positives, enhancing the signal-to-noise ratio. The Muse H2A.X Activation Dual Detection kit includes two directly conjugated antibodies-a phospho-specific anti-phospho-Histone H2A.X (Ser139)-Alexa Fluor® 555 and an anti-Histone H2A.X-PECy5 conjugated antibody-to measure total levels of Histone H2A.X. This two-color kit is designed to detect Histone H2A.X pathway activation by measuring H2A.X phosphorylation relative to the total H2A.X expression in any given cell population. By doing this, the levels of both the total and phosphorylated protein can be measured simultaneously in the same cell, resulting in a normalized and accurate measurement of H2A.X activation after stimulation.
All Muse Activation Dual Detection kits are optimized and designed for use on the Guava® Muse® cell analyzer. Both antibodies provided in the kit are carefully titrated and optimized together to ensure exceptional performance when run in multiplex, alleviating the need for any additional optimization. This kit contains optimized fixation, permeabilization, and assay buffers to provide researchers with a complete solution for H2A.X signaling analysis. Data generated using the Muse system provides statistical values, measuring the:
Percentage of inactivated cells
Percentage of activated cells (via phosphorylation)
Percentage of non-expressing cells