Description
Overview
The TREM-2 (Triggering Receptor Expressed on Myeloid Cells 2) family of receptors regulates the immune system's various cell types, such as neutrophils, monocyte/macrophages, microglia, and dendritic cells. TREM-2 requires the adaptor protein DAP12 for downstream signaling. Human cerebrospinal fluid (CSF) has been shown to include soluble TREM-2, which has been reported to be increased in people with multiple sclerosis and other inflammatory neurological illnesses compared to those without such conditions.
The AlphaLISA
SureFire
Human Total TREM2/DAP12 Complex is a sandwich immunoassay for the quantitative detection of total TREM2/DAP12 complex in cellular lysates, using Alpha technology.
Formats:
The HV (high volume) kit contains reagents to run 100 wells in 96-well format, using a 60 μL reaction volume.
The 500-point kit contains enough reagents to run 500 wells in 384-well format, using a 20 μL reaction volume.
The 10,000-point kit contains enough reagents to run 10,000 wells in 384-well format, using a 20 μL reaction volume.
The 50,000-point kit contains enough reagents to run 50,000 wells in 384-well format, using a 20 μL reaction volume.
AlphaLISA
SureFire Ultra
kits are compatible with:
Cell and tissue lysates
Antibody modulators
Biotherapeutic antibodies
Alpha
SureFire
kits can be used for:
Cellular kinase assays
Receptor activation studies
Screening
How it works
AlphaLISA
SureFire Ultra
TREM2/DAP12 complex assay principle
The AlphaLISA
SureFire Ultra
TREM2/DAP12 complex assay measures the TREM2/DAP12 complex level in a cell lysate.
The AlphaLISA
SureFire Ultra
TREM2/DAP12 complex assay uses two antibodies which recognize TREM2 and DAP12 proteins, respectively. AlphaLISA assays require two bead types: Acceptor and Donor beads. Acceptor beads are coated with a proprietary CaptSure™ agent to specifically immobilize the first assay antibody, labeled with a CaptSure™ tag. Donor beads are coated with streptavidin to capture the second antibody, which is biotinylated. In the presence of the TREM2/DAP12 complex, the two antibodies bring the Donor and Acceptor beads in close proximity whereby the singlet oxygen transfers energy to excite the Acceptor bead, enabling the generation of a luminescent Alpha signal. The amount of light emission is directly proportional to the quantity of TREM2/DAP12 complex present in the sample.
AlphaLISA
SureFire Ultra
TREM2/DAP12 complex two-plate assay protocol
The two-plate protocol involves culturing and treating the cells in a 96-well plate before lysis, then transferring lysates into a 384-well Optiplate plate before the addition of TREM2/DAP12 complex-AlphaLISA
SureFire Ultra
detection reagents. This protocol enables the cells viability and confluence to be monitored. In addition, lysates from a single well can be used to measure multiple targets.
AlphaLISA
SureFire Ultra
TREM2/DAP12 complex one-plate assay protocol
Detection of TREM2/DAP12 complex with AlphaLISA
SureFire Ultra
reagents can be performed in a single plate used for culturing, treatment, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust AlphaLISA
SureFire Ultra
quality.
