The Alpha SureFire® Ultra™ Multiplex kits allow the rapid, sensitive, and quantitative detection of two phosphoprotein targets in each well of an assay plate. This kit can be used in combination with any AlphaLISA SureFire Ultra assay to detect phospho-Ribosomal Protein S6 (Ser240/244) in a multiplexed assay.
For research use only. Not for use in diagnostic procedures. Please note this kit does not contain all buffers necessary to run the assay, and must be used with an AlphaLISA SureFire Ultra assay kit to obtain all required reagents.
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Alpha SureFire assays are cellular assays used to detect and quantitate phosphoproteins for cell signaling, pathway analysis, and cellular kinase studies using homogeneous (no wash steps; no separation steps) Alpha technology. Terbium SureFire assays enable multiplexing for detection of two phosphoproteins in a single well, when used in conjunction with any AlphaLISA SureFire Ultra kit. This multiplex measurement is achieved by the use of two types of Alpha Acceptor beads that emit at distinct wavelengths (Terbium: 545 nm, Europium: 615 nm). The two distinct Alpha Acceptor beads report their binding to distinct antigens through their association with specific assay antibodies.
The Terbium “Alpha 545 Acceptor Beads” in the kit are conjugated with a “CaptSure2” agent, which binds a specific CaptSure2 tag on one of the assay antibodies. In combination with the biotinylated antibody provided, the 545 nm signal generated will report levels of the phosphoprotein of interest (Target 2). This kit is to be used in conjunction with the reagents in an AlphaLISA SureFire Ultra kit, which will report that target-of-interest by Europium emission at 615 nm (Target 1).
*These assays utilize the bead-based Alpha Technology, and requires an Alpha Technology-compatible plate reader capable of reading dual emission wavelengths. Refer to the “AlphaPlex Quick Start Guide” and the “AlphaPlex Assay Development Guide” to find guidance about filters and mirrors selection, instrument protocol and channels crosstalk correction.
|Assay Target Class||Phosphoprotein|
|Product Brand Name||AlphaLISA SureFire Ultra Multiplex|
|Shipping Condition||Blue Ice|
|Unit Size||100 Assay Points|
The introduction of enzyme-linked immunosorbent assays (ELISAs) in the early 1970’s offered researchers a non-radiometric immunoassay platform without compromising sensitivity. Over the last 50 years scientists have made huge strides in disease research and drug discovery and a demand for greater assay throughput and sensitivity has evolved. In response, more robust immunoassays have been developed to address some of the limitations of the standard, colorimetric ELISA.
Find out about the most common limitations of traditional ELISAs and how different ELISA alternative technologies address these limitations.
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.