71784  EpiTag™ Cell Lysis Kit

This product has been discontinued.

The EpiTag™ product line has been discontinued. Most targets will be available in the MILLIPLEX EpiQuant portfolio in Q2 2012, with the exception of Total ERK1 and Total ERK2, which will be replaced with Total ERK1/2. To view the current offering from MILLIPLEX EpiQuant, please visit the website at : http://www.millipore.com/epiquant.

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WideScreen^® EpiTag™ assays utilize EpiTag™ Technology to detect protein- and phosphoprotein fragments rather than the traditional detection of intact proteins. Protein fragments are released by digestion with site-specific proteases and antibody pairs are generated to unique amino acid sequences, or EpiTag™ Sequences, on these fragments. This approach provides a high degree of specificity and the ability to multiplex assays for a protein’s total and phosphorylated levels without cross-reactivity. In addition, the assay can accurately measure concentrations of phosphorylated target proteins as synthetic phosphopeptides are used to generate standard curves.

The first WideScreen^® EpiTag assays have been developed to detect key signaling events in the ERK pathway from both human and mouse samples. ERK1 and ERK2 are serine/threonine kinases expressed broadly in tissues and various cell lines. ERKs are activated through their phosphorylation by the MAPK/ERK kinases MEK1 and MEK2. Once activated, ERK1 and ERK2 can phosphorylate many different proteins including cytoskeletal proteins, translation regulators, and transcription factors. ERK pathway protein levels and phosphorylation are deregulated in many types of cancer and are considered strong candidates for the development of therapeutic compounds.

The WideScreen^® EpiTag™ ERK Pathway Panel I Complete Kit contain all lysis, wash and digest buffers, premixed beads, and detection cocktails needed to run 96 tests. The components of the Complete Kits are also available separately, enabling greater options for cell culture and processing and the creation of custom multiplexes. To verify that various bead kits can be multiplexed together, consult consult the EpiTag Assay Compatibility Chart in TB499.

WideScreen^® EpiTag™ ERK Pathway Panel I Complete Kit Components
Premixed bead kits and detection antibodies for the multiplex quantitation of:

B-Raf ( pS446 )
MEK1/2 ( pS217, pS221/ pS222, pS226 )
ERK1 Total
ERK1 ( pT202/pY204 )
ERK2 Total ERK2 ( pT185/pY187 )

EpiTag™ Cell Lysis Kit
Lysis Reagent, Phosphatase Inhibitor Cocktail Set V, Benzonase^® Nuclease HC, Purity > 99%, 100X TCEP Solution, Aluminum Plate Sealer

EpiTag™ Digest Kit

a-Iodoacetamide, Endopeptidase Lys-C

WideScreen^® Reagent Kit
Streptavidin-PE Concentrate, 96-Well Filter Plate and Sealer, 5X Assay Diluent,10X Wash Buffer

Due to the unique nature of the EpiTag Technology and sample preparation, the EpiTag buffers and Bead Kits are incompatible with other WideScreen^® bead-based assay buffers and kits and those from other companies. EpiTag Technology and Assays are offered under limited, non-transferable license from Millipore Corporation.

 

The EpiTag™ Cell Lysis Kit contains a denaturing and reducing cell Lysis Reagent that efficiently releases proteins from the cell cytosol, membranes, and nucleus and is for use with the WideScreen^®. Benzonase^® Nuclease reduces sample viscosity caused by chromosomal DNA, and a cocktail of phosphatase inhibitors maintains the phosphorylation state of target proteins during cell lysis. An aluminum plate sealer is included for use when processing cells grown in 96-well plates. The kit is sufficient to make 5 ml of cell or tissue lysate (or 160 wells of cells grown in 96-well plates), and contains additional Lysis Reagent to prepare Titration Buffer for use in the WideScreen Assay.

Components

• Lysis Reagent
• Phosphatase Inhibitor Cocktail Set V (50X)
• Benzonase^® Nuclease HC, Purity > 99%
• 100X TCEP Solution
• Aluminum Plate Sealer

Due to the unique nature of the EpiTag Technology and sample preparation, the EpiTag buffers and Bead Kits are incompatible with other WideScreen^® brand bead-based assay buffers and kits and those from other companies. EpiTag Technology and Assays are offered under limited, non-transferable license from Millipore Corporation.