Apoptosis Assays

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Bio-Plex Pro RBM Apoptosis Assays - Bio-Plex Suspension Array

Overview

Available Bio-Plex Pro™ Apoptosis Assays
Apoptosis Panel 1 Apoptosis Panel 2 Apoptosis Panel 3
Analyte Bead Region Analyte Bead Region Analyte Bead Region
Bak 74 Bad 74 Active caspase-3 57
Bax 27 Bax/Bcl-2 dimer 42 Bcl-xL/Bak dimer 47
Lamin B, intact and
45 kD
14 Bcl-xL 22 Mcl-1/Bak dimer 54
Smac 19 Bim 12 Survivin 20
  Mcl-1 18  

 

Apoptosis Analyte Definitions
Description Apoptotic Action Apoptotic Cell
Bak Bcl-homologous antagonist/killer is a pro-apoptotic member of the Bcl-2 family. In healthy cells, it is integrated in the mitochondrial outer membrane. Upon apoptotic stimuli, Bak forms oligomer channels in the mitochondrial membrane for cytochrome C release. This activity is regulated by forming a complex with anti-apoptotic Mcl-1 and Bcl-xL. Pro-apoptotic Mitochondrial membrane
Bax Bcl-2 associated protein X is a pro-apoptotic member of the Bcl-2 family. In healthy cells, it is found as a monomer in the cytosol, but upon apoptotic stimuli, it translocates to the mitochondrial outer membrane and forms large oligomeric complexes. There it interacts with pore proteins to enable cytochrome C release into the cytosol and initiate the caspase activation pathway for apoptosis. Bax may cycle to the mitochondrial membrane and dimerize with Bcl-xL. Pro-apoptotic Moves to the mitochondrial membrane
Lamin B,
intact and 45 kD
Nuclear lamins are proteins of intermediate filament type located at the outer rim of the nucleus. They consist of two types of polypeptides, lamin A and lamin B. Lamin B consists of B1 and B2 subtypes. Lamins mechanically stabilize the cell nucleus and also play a role in DNA replication and chromatin organization. Lamin B is cleaved by caspase-3 and caspase-6 during the early phases of apoptosis (up to 90 min) before DNA fragmentation. Detection of cytoplasmic dissociated lamin B indicates cell apoptosis. Early indicator of apoptosis Cytosol
Smac Second mitochondria-derived activator of caspase is a dimeric mitochondrial protein synthesized in the cytoplasm as a 239 amino acid precursor protein, with 55 amino acids at the N-terminus serving as a mitochondrial-targeting sequence. Under apoptotic stimuli, it is proteolytically cleaved to a 23 kD active form and released into the cytosol together with cytochrome C, where it reverses IAP inhibition of caspase-9, allowing caspase-9 to activate the caspase cascade. Pro-apoptotic Cytosol
Bad Bcl-2-associated death promoter is a pro-apoptotic, BH3-only binding domain member of the Bcl-2 family. BH3-only proteins connect apoptotic death signals to the activation of Bax and Bak, which control mitochondrial membrane disruption and apoptosis. Phosphorylated BAD (pBAD) is typically bound to the cytosolic protein 14-3-3, and is thus sequestered away from the mitochondria. Dephosphorylation results in the release of cytosolic (free) BAD, which binds to and inhibits the pro-survival activity of Bcl-2 family proteins at the mitochondrial membrane. Pro-apoptotic Cytosol (free)
Bax/Bcl-2 dimer B-cell lymphoma-2 is an anti-apoptotic protein that resides on the outer mitochondrial membrane. When bound to Bax as a heterodimer, it inhibits permeability of the mitochondrial membrane, preventing release of cytochrome C. Anti-apoptotic Mitochondrial membrane; decreased with apoptosis
Bcl-xL B-cell lymphoma-extra large is an anti-apoptotic member of the Bcl-2 family found in the outer mitochondrial membrane. Heterodimerization with pro-apoptotic proteins (especially Bak) inhibits apoptosis by preventing release of cytochrome C. Anti-apoptotic Mitochondrial membrane
Bim Bcl-2-interacting mediator of cell death is a pro-apoptotic protein belonging to the BH3-only group of the Bcl-2 family. Bim binds and antagonizes pro-survival members of the Bcl-2 family such as Mcl-1, Bcl-xL, and Bcl-2. Three prominent isoforms are generated by alternative splicing: Bim-S, Bim-L, and Bim-EL. Each isoform has the ability to induce apoptosis. Pro-apoptotic Cytosol or mitochodrial membrane if dimerized
Mcl-1 Induced myeloid leukemia cell differentiation protein-1 is an anti-apoptotic member of the Bcl-2 family. Heterodimerization with pro-apoptotic proteins (especially Bak) inhibits apoptosis. While Mcl-1 may not be as potent a protector against apoptosis as Bcl-2, it does appear to be the main anti-apoptotic protein in some cell types, including neutrophils. Anti-apoptotic Mainly mitochondrial membrane, some in cytosol
Active Caspase-3 Cysteinyl aspartyl protease-3 belongs to the peptidase C14A enzyme family and is known to play an important role in the apoptotic cascade. The active enzyme is formed by cleavage of the inactive 32 kD pro-enzyme into the p17 and p12 subunits. Two of each subunit noncovalently heterodimerize, giving the final enzyme two catalytic sites. Active caspase-3 cleaves and activates other caspases and is a primary regulator of apoptotic-associated proteolysis. Pro-apoptotic Cytosol (high levels)
Bcl-xL/Bak dimer Anti-apoptotic Bcl-xL heterodimerizes with Bak at the mitochondrial outer membrane and inhibits permeability of the mitochondrial membrane, preventing release of cytochrome C. During apoptosis BH3-only proteins, such as Bim and Bad, will bind to Bcl-xL and cause Bak to be released. Upon release, Bak will oligomerize creating pores in the mitochondrial membrane. Anti-apoptotic Mitochondrial membrane; decreased with apoptosis
Mcl-1/Bak dimer Anti-apoptotic Mcl-1 heterodimerzes with Bax and Bak at the mitochondrial outer membrane to prevent their activation, thus inhibiting cytochrome C release from the mitochondria. Anti-apoptotic Mitochondria; decreased with apoptosis
Survivin Survivin is a 16 kD member of the Inhibitor of Apoptosis (IAP) family, which also plays a role in chromosome segregation and cytokinesis. The anti-apoptotic function of survivin comes from its ability to inhibit the activation of caspases-3 and -7. Survivin is found to be upregulated in various tumors. Anti-apoptotic Cytosol/nucleus
Apoptosis Signaling Pathway

Apoptosis Signaling Pathway

Apoptosis is induced by at least two distinct signaling pathways, extrinsic and intrinsic. The extrinsic pathway is triggered by signaling through death receptors such as Fas, followed by downstream activation of caspase-8 and caspase-3. The intrinsic pathway is triggered by cytotoxic stress, which leads to translocation of Bcl-2 family proteins, Bax and Bak, to the mitochondrial membrane. Oligomerization of Bax and Bak causes release of cytochrome C into the cytosol, which promotes apoptosome formation, caspase activation, degradation of nuclear Lamin B, and cell death. Pro-apoptotic proteins included in the panels (); Anti-apoptotic proteins included in the panels (); Proteins involved in apoptosis but not included in the panels ().

Use the Bio-Plex Pro RBM apoptosis multiplex assays for the detection of a highly relevant set of intracellular proteins involved in the commitment, onset, and induction of apoptosis by the intrinsic pathway.

 Panel 1 Analytes:
  • Bak
  • Bax
  • Lamin B
  • Smac
Panel 2 Analytes:
  • Bad
  • Bax/Bcl-2 dimer
  • Bcl-xL
  • Bim
  • Mcl-1
Panel 3 Analytes:
  • Active caspase-3
  • Bcl-xL/Bak dimer
  • Mcl-1/Bak dimer
  • Survivin

For use with cell lysate or tissue lysate samples.

Benefits of the Bio-Plex Pro RBM Multiplex Apoptosis Assays

  • Optimized for high precision and lot-to-lot reproducibility of sample measurements
  • Magnetic beads for simplified plate processing
  • 2-level quality controls with kit lot-specific ranges
  • Assay quick guide to get you started right away

Kit Components

  • 6.5 µm magnetic capture beads
  • Detection antibodies
  • Standards
  • 2-level controls
  • Standard diluent
  • Blocking buffer
  • Lysate dilution buffer (LDB)
  • Cytosolic extraction buffer (CEB)
  • 10x assay buffer
  • 10x streptavidin-PE
  • Flat bottom plate
  • Plate seals
  • Instructions

Instrument Compatibility

Compatible with these readers and wash stations:

More Information

To learn more about performance characteristics and other cancer-related panels, refer to the documents listed below:

Bio-Plex Pro™ RBM Apoptosis Panel 1

171-WAR1CK
1 x 96-well, includes magnetic capture beads, detection antibodies, standards, controls, standard diluent, buffers, streptavidin-PE, flat bottom plate, plate seals, for the detection of 4 apoptosis markers in cell and tissue lysates

List Price:   $1,430.00
Quantity:   Add to cart
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Bio-Plex Pro™ RBM Apoptosis Panel 2

171-WAR2CK
1 x 96-well, includes magnetic capture beads, detection antibodies, standards, controls, standard diluent, buffers, streptavidin-PE, flat bottom plate, plate seals, for the detection of 5 apoptosis markers in cell and tissue lysates

List Price:   $1,595.00
Quantity:   Add to cart
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Bio-Plex Pro™ RBM Apoptosis Panel 3

171-WAR3CK
1 x 96-well, includes magnetic capture beads, detection antibodies, standards, controls, standard dilluent, buffers, streptavidin-PE, flat bottom plate, plate seals, for the detection of 4 apoptosis markers in cell and tissue lysates

List Price:   $1,430.00
Quantity:   Add to cart
Add to Hot List   Add to Quote
Number Description Options
10033631 Instruction Manual, Bio-Plex Pro RBM Apoptosis Assays, Rev A Click to download
10033632 Instruction Manual, Bio-Plex Pro RBM Apoptosis Assays, Quick Guide, Rev B Click to download
6474 Bio-Plex Pro RBM Apoptosis Assays, Product Information Sheet, Rev A Click to download
6458 Bio-Plex Success Guide/Assay Sample Prep, Rev B Click to download
6518 Multiplex Analysis of Cell Death Markers Using Bio-Plex Pro RBM Apoptosis Assays, Rev A Click to download