Revision 1

#98863Store at -20C

1 Kit

(9 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
TREM2 (D8I4C) Rabbit mAb 91068 20 µl 28 kDa Rabbit IgG
TREM2 (E7P8J) Rabbit mAb (Carboxy-terminal Antigen) 76765 20 µl 11, 28 kDa Rabbit IgG
AMPKα (D5A2) Rabbit mAb 5831 20 µl 62 kDa Rabbit IgG
Phospho-AMPKα (Thr172) (40H9) Rabbit mAb 2535 20 µl 62 kDa Rabbit IgG
mTOR (7C10) Rabbit mAb 2983 20 µl 289 kDa Rabbit IgG
Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb 5536 20 µl 289 kDa Rabbit IgG
Akt (pan) (C67E7) Rabbit mAb 4691 20 µl 60 kDa Rabbit IgG
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb 4060 20 µl 60 kDa Rabbit IgG
LC3A/B (D3U4C) XP® Rabbit mAb 12741 20 µl 14, 16 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

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

Description

The TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit provides an economical means of detecting metabolic signaling pathways downstream of TREM2 by western blot. The kit includes enough antibodies to perform at least two western blot experiments with each primary antibody.

Storage

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.

Background

The triggering receptor expressed on myeloid cells 2 (TREM2) protein is an innate immune receptor that is expressed on the cell surface of microglia, macrophages, osteoclasts, and immature dendritic cells (1). The TREM2 protein plays a role in innate immunity and a rare functional variant (R47H) of TREM2 is associated with the late-onset risk of Alzheimer’s disease (AD)  (1,2). Research studies using mouse models of AD indicate that deficiency and haploinsufficiency of TREM2 can lead to increased β-amyloid (Aβ) accumulation as a result of dysfunctional microglia response (3). Activation of TREM2 in mouse models of AD ameliorates several forms of AD pathology, likely through a microglia-specific mechanism (4,5). This mechanism is under intense investigation, but may involve TREM2-dependent maintenance microglia energetic and biosynthetic metabolism (6). Autophagy is one mechanism by which cellular metabolism is maintained and, in the absence of TREM2, several AMPK-dependent autophagy cell signaling pathways are enhanced. AMP-activated protein kinase (AMPK) is highly conserved from yeast to plants and animals and plays a key role in the regulation of energy homeostasis (7). The tumor suppressor LKB1, in association with accessory proteins STRAD and MO25, phosphorylates AMPKα at Thr172 in the activation loop, and this phosphorylation is required for AMPK activation (8-10). AMPK is further regulated by several proteins within a regulatory cell signaling pathway. The mammalian target of rapamycin (mTOR, FRAP, RAFT) is a Ser/Thr protein kinase (11) that functions as an ATP and amino acid sensor to balance nutrient availability and cell growth (12). mTOR is phosphorylated at Ser2448 via the PI3 kinase/Akt signaling pathway and autophosphorylated at Ser2481 (13). Akt, also referred to as PKB or Rac, is regulated by phosphorylation at Ser473 (14,15). The presence of autophagy marker Light Chain 3 (LC3) in autophagosomes and the conversion of LC3 to the lower migrating form, LC3-II, have been used as indicators of autophagy (16).

  1. Colonna, M. (2003) Nat Rev Immunol 3, 445-53.
  2. Boutajangout, A. and Wisniewski, T. (2013) Int J Cell Biol 2013, 576383.
  3. Wang, Y. et al. (2015) Cell 160, 1061-71.
  4. Schlepckow, K. et al. (2020) EMBO Mol Med 12, e11227.
  5. Wang, S. et al. (2020) J Exp Med 217, e20200785.
  6. Ulland, T.K. et al. (2017) Cell 170, 649-663.e13.
  7. Hardie, D.G. (2004) J Cell Sci 117, 5479-87.
  8. Hawley, S.A. et al. (1996) J Biol Chem 271, 27879-87.
  9. Lizcano, J.M. et al. (2004) EMBO J 23, 833-43.
  10. Shaw, R.J. et al. (2004) Proc Natl Acad Sci U S A 101, 3329-35.
  11. Sabatini, D.M. et al. (1994) Cell 78, 35-43.
  12. Dennis, P.B. et al. (2001) Science 294, 1102-5.
  13. Navé, B.T. et al. (1999) Biochem J 344 Pt 2, 427-31.
  14. Burgering, B.M. and Coffer, P.J. (1995) Nature 376, 599-602.
  15. Franke, T.F. et al. (1995) Cell 81, 727-36.
  16. Kabeya, Y. et al. (2004) J Cell Sci 117, 2805-12.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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