Human TREM-2 kit HTRF®

This HTRF kit is designed for the rapid detection of human TREM-2 released by cells.

Ready-to-use
Highly specific
Faster and more convenient than ELISA

Download the protocols

This HTRF kit is designed for the rapid detection of human TREM-2 released by cells.

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Overview

The kit is designed for the rapid detection of TREM-2 (also known as Triggering Receptor Expressed on Myeloid Cells 2) in cell supernatant and whole cells. The TREM family of receptors regulates the activity of various cell types of the immune system, including neutrophils, monocyte/macrophages, microglia, and dendritic cells. Soluble TREM-2 has been detected in human cerebrospinal fluid (CSF), where it was found to be elevated in patients with multiple sclerosis and other inflammatory neurological conditions in comparison to patients without inflammatory neurologic disorders.

Benefits

CELLS AND CELL SUPERNATANT FORMATS
BIOMARKER FOR ALZHEIMER"S DISEASE

Assay principle

Human TREM2 is measured using a sandwich immunoassay involving two specific anti-human TREM2 antibodies, respectively labelled with Europium Cryptate (donor) and d2 (acceptor). The intensity of the signal is proportional to the concentration of the TREM2 present in the sample.

Assay Protocol

The simple TREM2 assay protocol, using a 384-well small volume white plate (20 µL final), is described on the right. Cell supernatant, sample, or standard is dispensed directly into the assay plate for the detection of human TREM2. The antibodies labelled with HTRF donor and acceptor may be pre-mixed and added in a single dispensing step to further streamline the assay procedure. The assay can be run in 96- to 384-well plates by simply resizing each addition volume proportionally.

TREM-2 detection on THP-1 cell supernatant

THP-1 cells were differentiated by adding 100 nM of PMA, causing the cells to become adherent. The final volume was 200µl in each well. The stimulation was done for 72 hours at 37°C. Cell supernatants from differentiated and un-differentiated conditions were then collected and tested in the assay. Several cell densities were assessed to ensure a correct concentration interpolation. THP-1 cells differentiated with PMA display a higher expression level of the TREM-2 receptor.

TREM2 detection on THP-1 cell supernantant

Soluble TREM2 quantification after washing treatments

Three different cellular densities of THP-1 cells were seeded in a 96-well plate and differentiated by adding 100nM of PMA, causing the cells to become adherent. The final volume was 200µl in each well. The stimulation was done for 72 hours at 37°C.

Cell supernatant was then collected for the soluble TREM2 quantification. Adherent cells were washed 3 times with complete cell culture medium, and the final supernatant was collected for soluble TREM2 detection. Cells were finally incubated for 24 hours at 37°C and a final sample was collected. For each sample, several dilutions were assessed to ensure a correct concentration interpolation.

The TREM2 receptor is highly expressed and cleaved after PMA induced THP-1 differentiation. The washing treatment enables the removal of remaining soluble TREM2. A 24 hour additional incubation enables the detection of newly released soluble TREM2.

Soluble TREM2 quantification upon washing treatments

Simplify pathway for TREM-2 (h) kit

TREM-2 is also known as Triggering Receptor Expressed on Myeloid Cells 2. The TREM family of receptors regulates the activity of various cell types in the immune system, including neutrophils, monocyte/macrophages, microglia, and dendritic cells. TREM-2 requires the adaptor protein DAP12 for downstream signaling. The extracellular part of TREM-2 can be cleaved, and this soluble TREM-2 has been detected in human cerebrospinal fluid (CSF). Soluble TREM-2 in patients with multiple sclerosis and other inflammatory neurological conditions has been found to be elevated compared to patients without inflammatory neurologic disorders.

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