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Alpha SureFire Ultra Multiplex phospho/total Cofilin Assay Kit, 50,000 Assay Points

The Alpha SureFire® Ultra™ Multiplex phospho Cofilin (Ser3) + Total Cofilin assay kit is used to measure both the phosphorylation (on Ser3) and total levels of endogenous Cofilin in cellular lysates. The assay is an ideal system for the screening of modulators of receptor activation (e.g. agonists and antagonists) as well as agents acting intracellularly, such as small molecule inhibitors of signal transduction. The assay will measure Cofilin phosphorylation by either recombinant or endogenous receptors and can be applied to primary cells.

For research use only. Not for use in diagnostic procedures.

Part Number
Unit Size
MPSU-PTCOF-K500
500 assay points
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MPSU-PTCOF-K10K
10,000 assay points
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MPSU-PTCOF-K50K
50,000 assay points
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MPSU-PTCOF-K-HV
100 assay points
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Overview

The Alpha SureFire Ultra Multiplex p-Cofilin (Ser3) + Total Cofilin assay kit is used to measure both phosphorylated and total levels of endogenous Cofilin in cellular lysates in a multiplexing Alpha no-wash assay. No specially engineered cell lines are required. The 615 nm (Eu) signal corresponds to the phosphorylated Cofilin analysis, and the 545 nm (Tb) signal corresponds to the total Cofilin analysis. This kit has been formulated to provide superior signal:background assay windows and to perform without interference in the presence of extraneous antibodies, making it amenable to the study of therapeutic and blocking antibodies.

  • No-wash steps, no separation steps
  • Multiplexing assay
  • Sensitive detection
  • No interference from exogenous antibodies
  • Results in less than 3 hours

In this assay, the Alpha 615 Acceptor bead is coated with the CaptSure™ antibody, which binds the CaptSure-tagged anti-phospho target antibody. The Alpha 545 Acceptor bead is coated with the CaptSure2™ agent, which binds the CaptSure2 tagged anti-total target protein antibody. The Alpha Donor bead binds the biotinylated anti-total target protein antibody.

Specifications

Assay Target Cofilin
Assay Target Class Phosphoprotein, Protein
Assay Technology Alpha
Automation Compatible Yes
Detection Method Alpha
Experimental Type In vitro
Product Brand Name AlphaLISA SureFire Ultra Multiplex
Shipping Condition Blue Ice
Therapeutic Area Central Nervous System
Unit Size 50,000 assay points
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Application Note

Alpha Terbium SureFire UltraMultiplex: Simultaneous Dual Phosphoprotein Target Analysis

Evaluating pharmaceutical compound efficacy and safety in regulating cell behavior can involve the study of multiple signal transduction pathways and measuring more than one cellular target can provide greater confidence of positive compound hits on the cellular target of interest. Protein phosphorylation has been identified as a useful readout of cellular activation or inhibition, and these pathways are commonly targeted for therapeutic modulation of disease.

Numerous assay technologies are available to look at protein phosphorylation for drug development, including the AlphaLISA SureFire Ultra which provides a highly sensitive, fully homogenous option for the analysis of signaling pathways. In this application note, we describe the implementation of multiplexing with the AlphaLISA SureFire Ultra Multiplex assay, the only multiplexing platform that is truly homogenous and high throughput.

PDF 1 MB
Simultaneous Detection of Total and Phosphorylated Protein in Cellular Kinase Assay Using Alpha SureFire Ultra Multiplex Technology

Cellular kinase signal transduction pathways are involved in the regulation of many important cellular processes such as cell survival, differentiation, and apoptosis. Kinase signaling networks are typically characterized by multiple kinases arranged in cascades containing nodes with feedback loops and crosstalk between pathways. Numerous assay technologies have been developed for studying kinase signaling pathways, and for screening compound libraries in search of agents to modify receptors or kinase activities. The quality of these assays can be impacted by data variability due to cell seeding inhomogeneity or from compound toxicity. A common method for controlling for variability is to normalize the assay signal to a cellular protein whose level does not change as a function of the treatment.

Alpha SureFire® Ultra Multiplex Phospho and Total assays utilize two types of Alpha Acceptor beads to simultaneously measure two signals from each assay well to easily normalize the assay. Here, we demonstrate the benefit and utility of normalizing assay signal of phosphorylated protein levels to total protein levels in two different cellular models: ERK 1/2 phosphorylation in human melanoma A375 cells and AKT 1/2/3 phosphorylation in mouse myoblast C2C12 cells.

PDF 1 MB

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Drug Discovery Screening Solutions Brochure

Find out about our range of integrated solutions for drug discovery screening in this e-brochure.

Our screening solutions for high-throughput screening, phenotypic screening and data analysis help to streamline drug discovery workflows in labs across the globe. Our portfolio includes automated liquid handling, assays and reagents, imaging and detection systems, and informatics.

Working independently or together, with our solutions you can achieve consistent and accurate results. By accelerating the identification and characterization of effective and safe drug candidates, the PerkinElmer portfolio enables you to optimize efficiency in your lab and deliver more actionable, real-world results.

Download the brochure to learn more about how we can partner with you so that you can discover smarter, more effective, data-driven breakthroughs in the critical screening stages of drug discovery and development.

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