Apoptosis is an essential physiological process that plays a crucial role in normal development and homeostatic mechanisms of multicellular organisms. At least two major apoptotic pathways have been described in mammalian cells: the intrinsic pathway via Caspase-9 activation, in which death arises from mitochondrial dysfunction, and the extrinsic pathway via Caspase-8 activation, in which death is initiated from the activation of cell surface receptors. Bcl-2 family members play an important role in the intrinsic apoptotic pathway and includes over 17 members. Interactions of these Bcl-2 family members function as a ‘life/death switch′ that integrates diverse inter- and intracellular cues to determine whether or not the stress apoptosis pathway should be activated. For example, Bcl-2 and its closely related anti-apoptotic proteins, Bcl-xL and Mcl-1, can sequester their pro-apoptotic counterparts (BAD, BIM, NOXA and PUMA) and thereby inhibit the initiation steps of apoptosis.The MILLIPLEX® MAP 4-plex Bcl-2 Family Apoptosis Panel 4 Magnetic Bead Kit (Cat. No. 48-683MAG) is a premixed kit to be used for the simultaneous quantification of the following 4 analytes:Bcl-xLMcl-1Bcl-2NOXA/Mcl-1
Cross-reactivity between the antibodies and any of the other analytes in this panel is non-detectable or negligible.
Intracellular Bead-Based Multiplex Assays using the Luminex technology enable the simultaneous relative quantitation of multiple phosphorylation and total pathway proteins in tissue and cell lysate samples. Compare Multiplexing results to those of Western blotting.An overnight (4°C) incubation is recommended for best results.This assay requires 25 μL diluted cell lysate per well.This kit must be run using Assay Buffer 1 (provided).The suggested working range of protein concentration for the assay is 1 to 25 μg of total protein/well (25 μL/well at 40 to 1,000 μg/mL).Analytes available:Bcl-xL;Mcl-1;BCL-2 (Total);NOXA/Mcl-1
Storage and Stability
Recommended storage for kit components is 2 - 8°C.
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MILLIPLEX is a registered trademark of Merck KGaA, Darmstadt, Germany
xMAP is a registered trademark of Luminex Corp
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