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98863
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit #98863

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TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 1
Western blot analysis of extracts from RD cells, untreated (-) or Torin 1-treated (250 nM, 4 hr; +), using LC3A/B (D3U4C) XP® Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 2
Western blot analysis of extracts from C2C12 cells, untreated or oligomycin-treated (0.5 µM), using Phospho-AMPKα (Thr172) (40H9) Rabbit mAb (upper) or AMPKα Antibody #2532 (lower).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 3
Western blot analysis of extracts from 293, A431, COS, C6, and C2C12 cells, using mTOR (7C10) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 4
Immunoprecipitation of mTOR protein from MCF-7 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is mTOR (7C10) Rabbit mAb. Western blot analysis was performed using mTOR (7C10) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 5
Flow cytometric analysis of A549 cells using mTOR (7C10) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 6
Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 7
Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from various cell lines, using Akt (pan) (C67E7) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 8
Western blot analysis of extracts from serum-starved NIH/3T3 cells, untreated or insulin-treated (150 nM, 5 minutes), alone or in combination with λ-phosphatase, using Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb (upper) or mTOR (7C10) Rabbit mAb #2983.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 9
Western blot analysis of extracts from HeLa, K-562, C6, and Neuro-2a cells using AMPKα (D5A2) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 10
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 11
Western blot analysis of extracts from mouse bone marrow derived macrophages (BMDM), untreated (-) or treated with peptide N-glycosidase F (PNGase F; +), and Neuro-2a cells using TREM2 (E7P8J) Rabbit mAb (Carboxy-terminal Antigen, Mouse Specific) (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 12
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc-tagged full-length human TREM2 protein (hTREM2-Myc; +), using TREM2 (D8I4C) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 13
Western blot analysis of extracts from HeLa, NIH/3T3, and KNRK cells, untreated (-) or chloroquine-treated (50 μM, overnight; +), using LC3A/B (D3U4C) XP® Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 14
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® mTOR siRNA II (+), using mTOR (7C10) Rabbit mAb #2983 and α-Tubulin (11H10) Rabbit mAb #2125. mTOR (7C10) Rabbit mAb confirms silencing of mTOR expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of mTOR siRNA.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 15
Immunoprecipitation of phospho-Akt (Ser473) from Jurkat extracts treated with Calyculin A #9902 (100nM, 30 min). Lane 1 is 10% input, lane 2 is Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb, and lane 3 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900. Western blot analysis was performed with Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded human melanoma using Akt (pan) (C67E7) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 17
Confocal immunofluorescent analysis of HeLa cells, rapamycin-treated (#9904, 10 nM for 2 hours, left), insulin-treated (150 nM for 6 minutes, middle) or insulin- and λ-phosphatase-treated (right), using Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 18
Western blot analysis of extracts from THP-1, HL-60, and Jurkat cells using TREM2 (D8I4C) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 19
Immunohistochemical analysis of paraffin-embedded mouse prostate using LC3A/B (D3U4C) XP® Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 20
Immunohistochemical analysis of paraffin-embedded NCI-H228 cell pellets, control (left) or phenformin-treated (right), using Phospho-AMPKalpha (T172) (40H9) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 21
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, showing cytoplasmic localization using mTOR (7C10) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 22
Immunohistochemical analysis of paraffin-embedded MDA-MB-468 xenograft using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (left) or PTEN (138G6) Rabbit mAb #9559 (right). Note the presence of P-Akt staining in the PTEN deficient MDA-MB-468 cells.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 23
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Akt (pan) (C67E7) Rabbit mAb in the presence of control peptide (left) or Akt (pan) Blocking Peptide #1085 (right).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 24
Immunoprecipitation of TREM2 from THP-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TREM2 (D8I4C) Rabbit mAb. Western blot analysis was performed using TREM2 (D8I4C) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 25
Immunohistochemical analysis of paraffin-embedded NIH/3T3 cell pellets, control (left) or chloroquine-treated (right), using LC3A/B (D3U4C) XP® Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 26
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-AMPKα (Thr172) (40H9) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 27
Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using mTOR (7C10) Rabbit mAb in the presence of control peptide (left) or mTOR Blocking Peptide #1072 (right).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 28
Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain® Antibody Diluent #8112 (left) to TBST/5% normal goat serum (right) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 29
Immunohistochemical analysis using Akt (pan) (C67E7) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 30
Confocal immunofluorescent analysis of THP-1 (positive, left) and HL-60 (negative, right) cells using TREM2 (D8I4C) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 31
Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using LC3A/B (D3U4C) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 32
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Phospho-AMPKα (Thr172) (40H9) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 33
Immunohistochemical analysis of paraffin-embedded mouse brain using mTOR (7C10) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 34
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 35
Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Akt (pan) (C67E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5 (fluorescent DNA dye).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 36
Confocal immunofluorescent analysis of HeLa (upper) and C2C12 (lower) cells, chloroquine-treated (50 μM, overnight; left), nutrient-starved with EBSS (3 hr, middle) or untreated (right) using LC3A/B (D3U4C) XP® Rabbit mAb (green) and β-Actin (13E5) Rabbit mAb (Alexa Fluor® 555 Conjugate) #8046 (red). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 37
Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Phospho-AMPKα (Thr172) (40H9) Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 38
Confocal immunofluorescent analysis of mouse embryonic fibroblast (MEF) cells using mTOR (7C10) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 39
Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb on SignalSlide® Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 40
Flow cytometric analysis of Jurkat cells using Akt (pan) (C67E7) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 41
Flow cytometric analysis of HeLa cells, untreated (blue) or treated with chloroquine (50 µM, 16 hr; green) using LC3A/B (D3U4C) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 42
Flow cytometric analysis of A549 cells using mTOR (7C10) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 43
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 44
Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb. (Tissue section courtesy of Dr. Sabina Signoretti, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.)
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 45
Immunohistochemical analysis of paraffin-embedded U-87MG xenograft, untreated (left) or lambda phosphatase-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 46
Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).
TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit: Image 47
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 2 hr; blue) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
To Purchase # 98863
製品# サイズ 数量 価格 在庫
98863T
1 Kit  (9 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
TREM2 (D8I4C) Rabbit mAb 91068 20 µl
  • WB
  • IP
  • IF
H 28 Rabbit IgG
TREM2 (E7P8J) Rabbit mAb (Carboxy-terminal Antigen, Mouse Specific) 76765 20 µl
  • WB
M 11, 28 Rabbit IgG
AMPKα (D5A2) Rabbit mAb 5831 20 µl
  • WB
  • IP
H M R Mk B 62 Rabbit 
Phospho-AMPKα (Thr172) (40H9) Rabbit mAb 2535 20 µl
  • WB
  • IP
  • IHC
H M R Hm Mk Dm Sc 62 Rabbit IgG
mTOR (7C10) Rabbit mAb 2983 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk 289 Rabbit IgG
Phospho-mTOR (Ser2448) (D9C2) XP® Rabbit mAb 5536 20 µl
  • WB
  • IP
  • IF
H M R Mk 289 Rabbit IgG
Akt (pan) (C67E7) Rabbit mAb 4691 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Mk Dm 60 Rabbit IgG
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb 4060 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R Hm Mk Dm Z B 60 Rabbit IgG
LC3A/B (D3U4C) XP® Rabbit mAb 12741 20 µl
  • WB
  • IHC
  • IF
  • F
H M R 14, 16 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

Product 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.

Specificity / Sensitivity

Each total antibody in the TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit detects endogenous levels of its target protein. TREM2 (D8I4C) Rabbit mAb and TREM2 (E7P8J) Rabbit mAb (Carboxy-terminal Antigen, Mouse Specific) detect both the full-length and the carboxy-terminal membrane fragment generated by proteolytic processing. TREM2 (E7P8J) Rabbit mAb (Carboxy-terminal Antigen, Mouse Specific) also detects a non-specific band of unknown origin migrating at ~80 kDa. LC3A/B (D3U4C) XP® Rabbit mAb recognizes endogenous levels of total LC3A and LC3B proteins. AMPKα (D5A2) Rabbit mAb detects both the α1 and α2 isoforms of the catalytic subunit. Each phospho-specific antibody in the TREM2-dependent mTOR Metabolic Fitness Antibody Sampler Kit detects endogenous levels of Akt only when phosphorylated at Ser473, AMPKα only when phosphorylated at Thr172, and mTOR protein only when phosphorylated at Ser2448. Phospho-AMPKα (Thr172) (40H9) Rabbit mAb detects both α1 and α2 isoforms of the catalytic subunit, but does not detect the regulatory β or γ subunits.

Source / Purification

Monoclonal antibodies to total proteins are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Leu221 of human TREM2 protein, Arg21 of human AMPKα protein, Ser2481 of human mTOR protein, Gly215 of mouse TREM2 protein, Leu44 of human LC3B protein (conserved in LC3A), and the carboxy-terminal sequence of mouse Akt protein. Phospho-specific monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Thr172 of human AMPKα protein, Ser2448 of human mTOR protein, and Ser473 of human Akt protein.

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.

Pathways & Proteins

Explore pathways + proteins related to this product.

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For Research Use Only. Not For Use In Diagnostic Procedures.
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U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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