Human TREM-1 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human TREM-1 Immunoassay is a 4.5 hour solid phase ELISA designed to measure TREM-1 levels in cell culture supernates, cell lysates, serum, plasma, and saliva. It contains NS0-expressed recombinant human TREM-1 and antibodies raised against the recombinant protein. Results obtained for naturally occurring human TREM-1 showed linear curves that were parallel to the standard curves obtained using the Quantikine Human TREM-1 Immunoassay standards. These results indicate that this kit can be used to determine relative mass values for natural human TREM-1.

Preparation and Storage

Background: TREM-1

The TREM family of receptors regulates the activity of various cell types of the immune system including neutrophils, monocyte/macrophages, microglia, and dendritic cells. The receptors are members of the immunoglobulin superfamily, characterized structurally by a single V-set immunoglobulin (Ig) domain and a short cytoplasmic tail. TREMs associate with the DAP12 signal adapter, an immunoreceptor tyrosine-based activation motif- (ITAM)-containing protein that is phosphorylated and transduces signals when activated.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.