The Muse^® PI3K 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 PI3K Activation Dual Detection kit includes two directly conjugated antibodies-a phospho-specific anti-phospho-Akt (Ser473), Alexa Fluor^® 555 and an anti-Akt, PECy5 conjugated antibody-to measure total levels of Akt. This two-color kit is designed to measure Akt phosphorylation relative to the total Akt 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 PI3K activation after stimulation. Moreover, simultaneous measurement of both total and phospho-Akt confirms target specificity of the phosphorylation event. Together, a total and phospho antibody duo performed in multiplex provides enhanced and more reliable detection of the phospho:total ratio within a mixed population.

All Muse Activation Dual Detection kits are optimized and designed for use on the Guava^® Muse^® cell analyzer. The antibody pair provided in the kit has been carefully titrated to ensure the ability to measure total and phospho-Akt simultaneously on the same protein for accurate determination of protein level and activation. This kit contains optimized fixation, permeabilization, and assay buffers to provide researchers with a complete solution for Akt 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

PRODUCT DETAILS

Application:        Flow Cytometry

Storage:              See Documentation

Data Sheets:      QRC 

EXAMPLE DATA

Recommended Staining Protocol