MILLIPLEX MAP Human Late Apoptosis Magnetic Bead 6-Plex Kit - Cell Signaling Multiplex Assay

MILLIPLEX MAP Human Early Apoptosis Magnetic Bead 6-Plex Kit, 48-669MAG

MILLIPLEX

1 kit

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. Cleaved and activated Caspase-8 or -9 can activate Caspase-3 and -7 which are responsible for the cleavage of various proteins leading to the biochemical and morphological features characteristic of apoptosis. Bcl-2 family members play a pivotal role in the regulation of apoptosis as they reside upstream of irreversible cellular damage and focus much of their efforts at the level of mitochondria. The BH3-only protein Bad promotes cell death by interacting with and inhibiting the pro-survival Bcl-2 proteins. Survival factors suppress Bad-mediated apoptosis by inducing phosphorylation of Bad by Akt. Phosphorylated Bad then binds to 14-3-3 instead of Bcl-2 or Bcl-XL, resulting in the liberation of the anti-apoptotic proteins and the promotion of cell survival. Irreversible cellular damage can also activate apoptosis by transcriptional activation of many pro-apoptotic genes by the transcription factors AP1 (the stress activated pathway, JNK) and p53 (DNA-damage response pathway).

This MILLIPLEX® MAP Human Early Apoptosis Magnetic Bead 6-Plex Kit is a Cell Signaling multiplex assay. The analytes available for this multiplex kit are: Akt (Ser473), BAD (Ser112), Bcl-2 (Ser70), Active Caspase 8 (Asp384), Active Caspase 9 (Asp315), JNK (Thr183/Tyr185), p53 (Ser46).

Multiplexing

• 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 2 (provided).
• 1 - 25 μg cell lysate/well (recommended starting concentration is 40 to 1,000 μg protein/mL)

Human

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Premixed

Cell Signaling

Luminex xMAP

96-well plate

MILLIPLEX MAP Human Early Apoptosis Magnetic Bead 6-Plex Kit, 48-669MAG

Cross-reactivity between the antibodies and any of the other analytes in this panel is non-detectable or negligible.

Active Caspase 8 (Asp384), Active Caspase 9 (Asp315), Akt (Ser473), BAD (Ser112), Bcl-2 (Ser70), JNK (Thr183/Tyr185), p53 (Ser46)