PhosphoPlus^® In-Cell Duets

PhosphoPlus^® In-Cell Duets (ICW Compatible) provide an easy means to assess protein activation status in whole cell assays using the LI-COR^® Biosciences Odyssey^® Infrared Imaging System. These kits contain an activation-state and total protein antibody to your target of interest, allowing levels of phosphorylated protein to be independently measured and normalized to total protein concentration. The antibodies were selected based on superior performance, and were validated to work in a multiplex assay format. A near-IR detection cocktail is also included.

• CST's highest quality XP^® antibodies directed against activation state targets are optimally paired with a total protein antibody, allowing for the best experimental outcome.
• Cocktails are pre-optimized and pre-tested to ensure there is no binding interference between phospho- and total protein antibodies, saving valuable time and reagents.
• Technical support is provided by our in-house High Content Screening group who developed the product and knows it best, which translates into a fast, thorough, and accurate response.

Analysis of MDA-MB-468 cells exposed to varying concentrations of LY294002 (PI3 Kinase Inhibitor) #9901 for 2 hours, followed by hEGF #8916 stimulation for 20 minutes. The phosphorylation status of Akt, as well as the total protein expression level, was measured simultaneously using the PhosphoPlus^® Akt (Ser473) In-Cell Duet (ICW Compatible) #7255. With increasing concentrations of LY294002, a significant decrease (~3-fold) in phospho-Akt signal as compared to the hEGF-stimulated control was observed, while total Akt protein levels remained the same and were used to normalize the data. When using phospho-Akt as a measurement, the IC₅₀ of this compound was 2.7 μM. Data and images were generated on the LI-COR^® Biosciences Odyssey^® Infrared Imaging System.

PhosphoPlus^® In-Cell Duets (ICW Compatible)

No.	Name	Applications	Reactivity
7263	PhosphoPlus® p44/42 MAPK (Erk1/2)(Thr202/Tyr204) In-Cell Duet (ICW Compatible)	ICW	H, M, R, Mk
7255	PhosphoPlus® Akt (Ser473) In-Cell Duet (ICW Compatible)	ICW	H, M, R, Mk

Applications Key: ICW=In-Cell Western 
Reactivity Key: H=Human  M=Mouse  R=Rat  Mk=Monkey