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PhosphoPlus® MLKL (Ser345) Antibody Duet
Primary Antibodies
Antibody Duet

PhosphoPlus® MLKL (Ser345) Antibody Duet #24653

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other Image 1 - PhosphoPlus® MLKL (Ser345) Antibody Duet

Confocal immunofluorescent analysis of L-929 cells, untreated (left), pre-treated with Z-VAD (20 μM, 30 min) followed by treatment with SM-164 (100 nM) and Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/mL, 2.5 hr; center) and then post-processed with λ-phosphatase (right), using Phospho-MLKL (Ser345) (D6E3G) Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087 (fluorescent DNA dye).

other Image 2 - PhosphoPlus® MLKL (Ser345) Antibody Duet

Immunoprecipitation of MLKL protein from BaF3 cells. Lane 1 is 10% input, lane 2 is immunoprecipitated with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is MLKL (D6W1K) Rabbit mAb (Mouse Specific). Western blot was performed with MLKL (D6W1K) Rabbit mAb (Mouse Specific).

other Image 3 - PhosphoPlus® MLKL (Ser345) Antibody Duet

Western blot analysis of L-929 cells, untreated (-), or treated with combinations of the following treatments as indicated: Z-VAD (20 μM, added 30 min prior to other compounds; +), Mouse Tumor Necrosis Factor-α (mTNF-α) #5178 (20 ng/ml, 4 hr; +), SM-164 (100 nM, 4 hr; +), and necrostatin-1 (Nec-1, 50 μM, 4 hr; +), using Phospho-MLKL (Ser345) (D6E3G) Rabbit mAb (upper), total MLKL (D6W1K) Rabbit mAb (Mouse Specific) #37705 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower).

other Image 4 - PhosphoPlus® MLKL (Ser345) Antibody Duet

Western blot analysis of extracts from various cell lines using MLKL (D6W1K) Rabbit mAb (Mouse Specific).

To Purchase # 24653S
Product # Size Price
1 Kit N/A

Product Includes Quantity Reactivity MW(kDa) Isotype
Phospho-MLKL (Ser345) (D6E3G) Rabbit mAb 37333 100 µl M 54 Rabbit IgG
MLKL (D6W1K) Rabbit mAb (Mouse Specific) 37705 100 µl M 54 Rabbit IgG

Kit Usage Information


Product Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.


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 (1,2). The process is negatively regulated by caspases and is initiated through a complex containing the RIP1 and RIP3 kinases, typically referred to as the necrosome. Mixed lineage kinase domain-like protein (MLKL) is a pseudokinase that was identified as downstream target of RIP3 in the necroptosis pathway (3,4). During necroptosis RIP3 is phosphorylated at Ser227, which recruits MLKL and leads to its phosphorylation at Thr357 and Ser358 (3). Knockdown of MLKL through multiple mechanisms results in inhibition of necroptosis (3-5). While the precise mechanism for MLKL-induced necroptosis is unclear, some studies have shown that necroptosis leads to oligomerization of MLKL and translocation to the plasma membrane, where it effects membrane integrity (6-9).

  1. Christofferson, D.E. and Yuan, J. (2010) Curr Opin Cell Biol 22, 263-8.
  2. Kaczmarek, A. et al. (2013) Immunity 38, 209-23.
  3. Sun, L. et al. (2012) Cell 148, 213-27.
  4. Wang, Z. et al. (2012) Cell 148, 228-43.
  5. Wu, J. et al. (2013) Cell Res 23, 994-1006.
  6. Cai, Z. et al. (2014) Nat Cell Biol 16, 55-65.
  7. Chen, X. et al. (2014) Cell Res 24, 105-21.
  8. Wang, H. et al. (2014) Mol Cell 54, 133-46.
  9. Dondelinger, Y. et al. (2014) Cell Rep 7, 971-81.

Pathways & Proteins

Explore pathways + proteins related to this product.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus is a trademark of Cell Signaling Technology, Inc.
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