Revision 1

#73966Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
RIP (E8S7U) XP® Rabbit mAb 73271 20 µl 78 kDa Rabbit IgG
Phospho-RIP (Ser166) (D1L3S) Rabbit mAb 65746 20 µl 78-82 kDa Rabbit IgG
RIP3 (E7A7F) XP® Rabbit mAb 10188 20 µl 46-62 kDa Rabbit IgG
Phospho-RIP3 (Ser227) (D6W2T) Rabbit mAb 93654 20 µl 46-62 kDa Rabbit IgG
MLKL (E7V4W) Mouse mAb 26539 20 µl 54 kDa Mouse IgG2b kappa
Phospho-MLKL (Ser358) (D6H3V) Rabbit mAb 91689 20 µl 54 kDa Rabbit IgG
Cleaved Caspase-8 (Asp374) (E6H8S) Rabbit mAb 98134 20 µl 18, 41, 43 kDa Rabbit IgG
Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb 9664 20 µl 17, 19 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.


The Apoptosis/Necroptosis Antibody Sampler Kit II provides an economical means of detecting markers for apoptosis and necroptosis. The kit contains enough primary antibody to perform at least two western blot experiments.


Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/mL BSA, 50% glycerol, and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.


Apoptosis is a regulated physiological process leading to cell death (1,2). Caspases, a family of cysteine acid proteases, are central regulators of apoptosis. Caspases are synthesized as inactive zymogens containing a pro-domain followed by large (p20) and small subunits (p10) that are proteolytically processed in a cascade of caspase activity. Initiator caspases (including 8, 9, 10, and 12) are closely coupled to proapoptotic signals. Once activated, these caspases cleave and activate downstream effector caspases (including 3, 6, and 7), which in turn cleave cytoskeletal and nuclear proteins like PARP, α-fodrin, DFF, and lamin A, and induce apoptosis. Cytochrome c released from mitochondria is coupled to the activation of caspase-9, a key initiator caspase. Apoptosis induced through the extrinsic mechanisms involving death receptors in the tumor necrosis factor receptor superfamily activates caspase-8. Activated caspase-8 cleaves and activates downstream effector caspases, such as caspase-1, -3, -6, and -7. Caspase-3 is a critical executioner of apoptosis, as it is either partially or totally responsible for the proteolytic cleavage of many key proteins, such as the nuclear enzyme poly (ADP-ribose) polymerase (PARP).

Necroptosis, a regulated pathway for necrotic cell death, is triggered by a number of inflammatory signals, including cytokines in the tumor necrosis factor (TNF) family, pathogen sensors such as toll-like receptors (TLRs), and ischemic injury (3,4). Necroptosis is negatively regulated by caspase-8 mediated apoptosis in which the kinase RIP/RIPK1 is cleaved (5). Furthermore, necroptosis is inhibited by a small molecule inhibitor of RIP, necrostatin-1 (Nec-1) (6). Research studies show that necroptosis contributes to a number of pathological conditions, and Nec-1 has been shown to provide neuroprotection in models such as ischemic brain injury (7). RIP is phosphorylated at several sites within the kinase domain that are sensitive to Nec-1, including Ser14, Ser15, Ser161, and Ser166 (8). Phosphorylation drives association with RIP3, which is required for necroptosis (9-11). Mixed lineage kinase domain-like protein (MLKL) is a pseudokinase that was identified as a downstream target of RIP3 in the necroptosis pathway (12). During necroptosis, RIP3 is phosphorylated at Ser227, which recruits MLKL and leads to its phosphorylation at Thr357 and Ser358 (12). Knockdown of MLKL through multiple mechanisms results in inhibition of necroptosis (13). Phosphorylation of MLKL during necroptosis leads to its oligomerization with pore formation that affects membrane integrity (14-17).

  1. Degterev, A. et al. (2003) Oncogene 22, 8543-67.
  2. Green, D.R. (1998) Cell 94, 695-8.
  3. Christofferson, D.E. and Yuan, J. (2010) Curr Opin Cell Biol 22, 263-8.
  4. Kaczmarek, A. et al. (2013) Immunity 38, 209-23.
  5. Lin, Y. et al. (1999) Genes Dev 13, 2514-26.
  6. Degterev, A. et al. (2008) Nat Chem Biol 4, 313-21.
  7. Degterev, A. et al. (2005) Nat Chem Biol 1, 112-9.
  8. Ofengeim, D. and Yuan, J. (2013) Nat Rev Mol Cell Biol 14, 727-36.
  9. Cho, Y.S. et al. (2009) Cell 137, 1112-23.
  10. He, S. et al. (2009) Cell 137, 1100-11.
  11. Zhang, D.W. et al. (2009) Science 325, 332-6.
  12. Sun, L. et al. (2012) Cell 148, 213-27.
  13. Wu, J. et al. (2013) Cell Res 23, 994-1006.
  14. Cai, Z. et al. (2014) Nat Cell Biol 16, 55-65.
  15. Chen, X. et al. (2014) Cell Res 24, 105-21.
  16. Wang, H. et al. (2014) Mol Cell 54, 133-46.
  17. Dondelinger, Y. et al. (2014) Cell Rep 7, 971-81.

Background References

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