Human TREM2 Antibody

R&D Systems | Catalog # AF1828

R&D Systems
Discontinued Product
AF1828 has been discontinued. View all TREM2 products.

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Western Blot, ELISA, Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, Bioassay, ELISA Capture, ELISA Development (Capture), Functional Assay, Stimulation

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all TREM2 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant human TREM2
His19-Ser174
Accession # Q9NZC2

Specificity

Detects human TREM2 in direct ELISAs and Western blots. In direct ELISAs, less than 50% cross-reactivity with recombinant mouse TREM2b is observed.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human TREM2 Antibody

Detection of Human TREM2 antibody by Western Blot.

Detection of Human TREM2 by Western Blot.

Western blot shows lysates of THP-1 human acute monocytic leukemia cell line. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Human TREM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1828) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for TREM2 at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

TREM2 antibody in Human Dendritic Cells by Immunocytochemistry (ICC).

TREM2 in Human Dendritic Cells.

TREM2 was detected in immersion fixed immature human dendritic cells using Goat Anti-Human TREM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1828) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Human TREM2 Antibody in ELISA Standard Curve.

Human TREM2 ELISA Standard Curve.

Recombinant Human TREM2 protein was serially diluted 2-fold and captured by Mouse Anti-Human TREM2 Monoclonal Antibody(Catalog # MAB18281) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human TREM2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1828) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998) followed by Substrate Solution (Catalog # DY999) and stopping the enzymatic reaction with Stop Solution (Catalog # DY994).

Detection of Human TREM2 by Western Blot

Detection of Human TREM2 by Western Blot

TREM2 is shed at a single cleavage site in the ectodomain between histidine 157 and serine 158Regulated intramembrane proteolysis of TREM2. ADAM10 initiates proteolytic processing of TREM2 by liberating its ectodomain (sTREM2). Subsequent processing of the membrane retained C‐terminal fragment (CTF) by gamma ‐secretase within the transmembrane domain (TM) releases the intracellular domain (ICD) into the cytosol. A short p3‐like peptide (Haass et al, 1993) may be secreted.Outline of strategy I for mass spectrometric (MS) determination of the N‐terminal end of the TREM2 CTF enriched upon gamma ‐secretase inhibition using DAPT.Western blot analysis of TREM2 stably expressed in HEK293 Flp‐In cells upon gamma ‐secretase inhibition using DAPT. Application of DAPT leads to a robust accumulation of the TREM2 CTF under constitutive conditions as well as upon phorbol 12‐myristate 13‐acetate (PMA) mediated stimulation of TREM2 ectodomain shedding. The TREM2 9D11 antibody raised against the TREM2 C‐terminus was used to detect TREM2, and calnexin levels were analyzed as a loading control. Here, as in all other Western blots in Figs 1 and 2, the two lanes represent samples from two separate wells seeded at the same time.MALDI‐TOF MS determination of the ectodomain cleavage site by immunoprecipitation of TREM2 CTF. The peak at 8,841.48 Da corresponds to a single cleavage site between histidine 157 and serine 158. Very minor additional peaks may represent cellular degradation products, as the N‐terminal counterpart cannot be observed (see Fig 1I).Stimulation of ectodomain shedding by PMA leads to strong increases in sTREM2 (anti‐HA, upper panel) and sAPP alpha (2D8, lower panel). DMSO served as a vehicle control. EV, empty vector control.MALDI‐TOF MS determination of the ectodomain cleavage site upon ADAM17 stimulation using PMA. TREM2 CTFs were enriched by gamma ‐secretase inhibition using DAPT (Fig 1C). The peak at 8,837.09 Da corresponds to a single cleavage site between histidine 157 and serine 158.Outline of strategy II for MS determination of the C‐terminal end of sTREM2 generated by ectodomain shedding.Western blot analysis of full‐length TREM2‐TEVFlag (middle panel) and ectodomain shedding (upper panel) upon transient transfection of HEK293 Flp‐In cells. EV, empty vector control.MALDI‐TOF MS determination of the ectodomain cleavage site following the strategy outlined in Fig 1G. The peak at a mass of 3,947.78 Da corresponds to cleavage C‐terminal of histidine 157 and is absent in cells transfected with an empty expression vector.TREM2 is shed predominantly C‐terminal to histidine 157 as observed in two different cell types as well as under constitutive and PMA‐stimulated conditions.Western blot analysis of full‐length TREM2‐TEVFlag (middle panel) and ectodomain shedding (upper panel) upon transient transfection of THP‐1 monocytes.MALDI‐TOF MS determination of the ectodomain cleavage site in THP‐1 monocytes following the strategy outlined in Fig 1G. The peak at a mass of 3,943.80 Da corresponds to cleavage C‐terminal of histidine 157 and is absent in cells transfected with an empty expression vector.Source data are available online for this figure. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28855300), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human TREM2 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

ELISA

This antibody functions as an ELISA detection antibody when paired with Mouse Anti-Human TREM2 Monoclonal Antibody.

This product is intended for assay development on various assay platforms requiring antibody pairs. We recommend the Human TREM2 DuoSet ELISA Kit (Catalog # DY1828-05) for convenient development of a sandwich ELISA.

Flow Cytometry

2.5 µg/106 cells
Sample: Human peripheral blood monocytes

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed THP‑1 human acute monocytic leukemia cell line and immersion fixed immature human dendritic cells

Western Blot

0.2 µg/mL
Sample: THP‑1 human acute monocytic leukemia cell line

Reviewed Applications

Read 4 reviews rated 4.8 using AF1828 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: TREM2

TREM2 (Triggering Receptor Expressed on Myeloid cells-2) is a 35 kDa molecular weight type I transmembrane member of the TREM family and Ig superfamily. Mature human TREM2 consists of a 156 amino acid (aa) extracellular domain (ECD) with one V-type Ig-like domain, a 21 aa transmembrane (TM) domain, and a 35 aa cytoplasmic tail. Within the ECD, human TREM2 shares 73% and 74% aa sequence identity with mouse and rat TREM2, respectively. Two closely related transcripts were reported in mouse and designated TREM2a and TREM2b. Soluble forms of the TREM2 ECD are generated by alternative splicing or proteolytic cleavage, and the cytoplasmic domain can be liberated by gamma-Secretase mediated intramembrane cleavage. It is a pattern recognition receptor that binds anionic ligands. A positively charged lysine within the transmembrane segment allows association with the signal adapter protein, DAP12 to deliver an activating signal that plays a role in both innate and adaptive immune responses, including inhibition of macrophage activation. TREM2 is expressed on macrophages, immature myeloid dendritic cells, osteoclasts, microglia, and adipocytes. It promotes the differentiation and function of osteoclasts, the production of inflammatory cytokines by adipocytes, insulin resistance, and the phagocytic clearance of bacteria. In the CNS, TREM2 binds to ApoE, ApoA1, and ApoB and mediates the clearance of apoptotic neurons, amyloid plaques, and cell debris following demyelination. TREM2 also interacts with and modifies signaling through Plexin A1 on dendritic cells and osteoclasts. Mutations in TREM2 or DAP12 are associated with the development of Alzheimer's disease and Nasu-Hakola disease (NHD/PLOSL) which is characterized by presenile dementia and bone cysts. Soluble TREM2 is elevated in cerebrospinal fluid of patients with active multiple sclerosis (MS), and TREM2 blockade exacerbates disease symptoms in the experimental EAE model of MS.

Long Name

Triggering Receptor Expressed on Myeloid Cells 2

Alternate Names

PLOSL2, TREM-2

Entrez Gene IDs

54209 (Human); 102133279 (Cynomolgus Monkey)

Gene Symbol

TREM2

UniProt

Q9NZC2

Additional TREM2 Products

Product Documents for Human TREM2 Antibody

Certificate of Analysis

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Product Specific Notices for Human TREM2 Antibody

For research use only

Citations for Human TREM2 Antibody

Customer Reviews for Human TREM2 Antibody (4)

4.8 out of 5
4 Customer Ratings
5 Stars
75%
4 Stars
25%
3 Stars
0%
2 Stars
0%
1 Stars
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Human TREM2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 06/07/2023
    HEK293T cell stably expressing human TREM2
    Human TREM2 Antibody AF1828
  • Human TREM2 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Melanoma tissue
    Species: Human
    Verified Customer | Posted 11/04/2021
    Human TREM2 Antibody AF1828
  • Human TREM-2 Antibody
    Name: Dennis Berner
    Application: Western Blot
    Sample Tested: HEK293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 06/08/2017
    Human TREM2 Antibody AF1828
  • Human TREM-2 Antibody
    Name: Ju Shi
    Application: Flow Cytometry
    Sample Tested: 293T human embryonic kidney cell line
    Species: Human
    Verified Customer | Posted 05/16/2017

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