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98240
Mouse TREM2 Activity Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Mouse TREM2 Activity Antibody Sampler Kit #98240

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Mouse TREM2 Activity Antibody Sampler Kit: Image 1
Confocal immunofluorescent analysis of SW620 cells (left, positive) and ACHN cells (right, negative) using Syk (D3Z1E) XP® Rabbit mAb (green), DyLight 650 Phalloidin #12956 (red), and DAPI #4083 (blue).
Mouse TREM2 Activity Antibody Sampler Kit: Image 2
Western blot analysis of extracts from THP-1, U-937, and Raw 264.7 cells using DAP12 (D7G1X) Rabbit mAb.
Mouse TREM2 Activity Antibody Sampler Kit: Image 3
Western blot analysis of extracts from various cell lines using Syk (D3Z1E) XP® Rabbit mAb.
Mouse TREM2 Activity Antibody Sampler Kit: Image 4
Western blot analysis of extracts from Jurkat cells treated with hydrogen peroxide (2mM for 2 minutes) or with lambda phosphatase and extracts from Ramos cells treated with anti-human IgM (12 micrograms/ml for 2 minutes) using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb.
Mouse TREM2 Activity Antibody Sampler Kit: Image 5
Western blot analysis of extracts from mouse bone marrow derived macrophages (BMDM) cells, untreated (-) or treated with peptide N-glycosidase F (PNGase F; +), and Neuro-2a cells using TREM2 (E6T1P) Rabbit mAb (Amino-terminal Antigen, Mouse Specific) (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Mouse TREM2 Activity Antibody Sampler Kit: Image 6
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.
Mouse TREM2 Activity Antibody Sampler Kit: Image 7
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).
Mouse TREM2 Activity Antibody Sampler Kit: Image 8
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human DAP12 (hDAP12; +), using DAP12 (D7G1X) Rabbit mAb.
Mouse TREM2 Activity Antibody Sampler Kit: Image 9
Immunoprecipitation of Syk protein from SR cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Syk(D3Z1E) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Syk (D3Z1E) XP® Rabbit mAb
Mouse TREM2 Activity Antibody Sampler Kit: Image 10
Western blot analysis of extracts from THP-1 WT (left) or SYK KO (right) using Syk (D3Z1E) XP® Rabbit mAb (upper). Membranes stained with Ponceau S for total protein normalization (lower).  These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
Mouse TREM2 Activity Antibody Sampler Kit: Image 11
Confocal immunofluorescent analysis of Ramos cells, human IgM treated (left) or untreated (right), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb (green). Red = Propidium Iodide (PI)/RNase Staining Solution #4087.
Mouse TREM2 Activity Antibody Sampler Kit: Image 12
Immunoprecipitation of DAP12 from THP-1 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or DAP12 (D7G1X) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using DAP12 (D7G1X) Rabbit mAb.
Mouse TREM2 Activity Antibody Sampler Kit: Image 13
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Syk (D3Z1E) XP® Rabbit mAb.
Mouse TREM2 Activity Antibody Sampler Kit: Image 14
Flow cytometric analysis of Ramos cells, untreated (blue) or treated with anti-IgM (12 µg/mL, 2 min; green), using Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) 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.
Mouse TREM2 Activity Antibody Sampler Kit: Image 15
Immunohistochemical analysis of paraffin-embedded human lymph node using Syk (D3Z1E) XP® Rabbit mAb.
Mouse TREM2 Activity Antibody Sampler Kit: Image 16
Immunohistochemical analysis of paraffin-embedded mouse spleen using Syk (D3Z1E) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Mouse TREM2 Activity Antibody Sampler Kit: Image 17
Flow cytometric analysis of RL cells using Syk (D3Z1E) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.
To Purchase # 98240
Cat. # Size Qty. Price
98240T
1 Kit  (5 x 20 microliters)
$ 455

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
TREM2 (E7P8J) Rabbit mAb (Carboxy-terminal Antigen, Mouse Specific) 76765 20 µl
  • WB
M 11, 28 Rabbit IgG
TREM2 (E6T1P) Rabbit mAb (Amino-terminal Antigen, Mouse Specific) 61788 20 µl
  • WB
M 28 Rabbit IgG
Syk (D3Z1E) XP® Rabbit mAb 13198 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 72 Rabbit IgG
Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb 2717 20 µl
  • WB
  • IF
  • F
H M 70, 72 Rabbit IgG
DAP12 (D7G1X) Rabbit mAb 12492 20 µl
  • WB
  • IP
H M 10, 12 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The Mouse TREM2 Activity Antibody Sampler Kit provides an economical means of evaluating key members of the mouse TREM2 signaling pathway using phospho-specific and control antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody. 


Specificity / Sensitivity

Each antibody in the Mouse TREM2 Activity Antibody Sampler Kit detects endogenous levels of its target protein. TREM2 (E7P8J) Rabbit mAb (Carboxy-terminal Antigen, Mouse Specific) recognizes endogenous levels of total mouse TREM2 protein, both the full-length and the carboxy-terminal membrane fragment generated by proteolytic processing. A non-specific band of unknown origin is observed migrating at ~80 kDa. TREM2 (E6T1P) Rabbit mAb (Amino-terminal Antigen, Mouse Specific) recognizes endogenous levels of total TREM2 protein. A non-specific band of unknown origin is observed migrating at ~75 kDa. Syk (D3Z1E) XP® Rabbit mAb recognizes endogenous levels of total Syk protein. Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb detects endogenous levels of Zap-70 only when phosphorylated at Tyr319. It cross-reacts with endogenous levels of Syk when phosphorylated at Tyr352. DAP12 (D7G1X) Rabbit mAb recognizes endogenous levels of total DAP12 protein.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to residues surrounding Gly215 of mouse TREM2 protein, near the amino terminus of mouse TREM2 protein, Asn463 of human Syk protein, Tyr319 of human Zap-70, and near the carboxy terminus of human DAP12 protein.

Background

Alzheimer's Disease (AD) is one of the most common neurodegenerative diseases worldwide. Clinically, it is characterized by the presence of extracellular amyloid plaques and intracellular neurofibrillary tangles, resulting in neuronal dysfunction and cell death. Triggering receptor expressed on myeloid cells 2 (TREM2), a protein localized at the membrane of innate immune cells, including microglia in the brain, has been genetically linked to AD, with specific variants increasing disease risk by as much as threefold (1,2). The TREM2 receptor is a single-pass type I membrane glycoprotein that consists of an extracellular immunoglobulin-like domain, a transmembrane domain, and a cytoplasmic tail. Upon activation, TREM2 interacts with the tyrosine kinase-binding protein DNAX-activating protein 12 (DAP12, TYROBP) to form a receptor-signaling complex. The DAP12 protein structure consists of a short extracellular domain, a transmembrane domain, and a cytoplasmic immunoreceptor tyrosine-based activation motif (ITAM) (2-9). ITAMs function as a binding site for tyrosine kinases, including spleen tyrosine kinase (Syk). Syk is comprised of two tandem amino-terminal Src homology (SH) 2 domains separated by an SH2-kinase linker, and a C-terminal tyrosine kinase domain, separated from the SH2 domains by an inter-domain linker. When Syk binds to an ITAM, it changes conformation, allowing for residues within the inter-domain linker region, including Tyr352, to become phosphorylated. Residues within the activation loop subsequently become phosphorylated, leading to full Syk activation. Tyr525 and Tyr526 are located in the activation loop of the Syk kinase domain and phosphorylation at these residues (equivalent to Tyr519/520 of mouse Syk) is essential for Syk function (10-12). This activation can lead to the mediation of a variety of cellular responses, including proliferation, differentiation, inflammation, and phagocytosis. Evidence suggests that TREM2 and DAP12 may act in a Syk-dependent manner to drive microglial cellular responses in AD (2,4-8,13).
  1. Nguyen, A.T. et al. (2020) Acta Neuropathol 140, 477-493.
  2. Gratuze, M. et al. (2018) Mol Neurodegener 13, 66.
  3. Jonsson, T. et al. (2013) N Engl J Med 368, 107-16.
  4. Jay, T.R. et al. (2017) Mol Neurodegener 12, 56.
  5. McQuade, A. et al. (2020) Nat Commun 11, 5370.
  6. Schlepckow, K. et al. (2020) EMBO Mol Med 12, e11227.
  7. Zhao, Y. et al. (2018) Neuron 97, 1023-1031.e7.
  8. Colonna, M. (2003) Nat Rev Immunol 3, 445-53.
  9. Lanier, L.L. et al. (1998) Nature 391, 703-7.
  10. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7.
  11. Mansueto, M.S. et al. (2019) J Biol Chem 294, 7658-7668.
  12. Grädler, U. et al. (2013) J Mol Biol 425, 309-33.
  13. Turner, M. et al. (2000) Immunol Today 21, 148-54.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
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