Human Pentraxin 3/TSG-14 Quantikine ELISA Kit Summary

Product Summary

The Quantikine^® Human Pentraxin 3/TSG-14 Immunoassay is a 5.0 hour solid-phase ELISA designed to measure human Pentraxin 3 in serum-free cell culture supernates, plasma, and saliva. It contains NS0-expressed recombinant human Pentraxin 3 and antibodies raised against the recombinant protein. Natural human Pentraxin 3 showed linear curves that were parallel to the standard curves obtained using the Quantikine^® kit standards. These results indicate that this kit can be used to determine relative mass values for natural human Pentraxin 3.

Preparation and Storage

Background: Pentraxin 3/TSG-14

Pentraxin 3 (PTX3), also known as TNF-stimulated gene 14 (TSG-14), is a secreted glycoprotein belonging to the Pentraxin family of proteins. It is the prototypical long Pentraxin, exhibiting a C-terminal Pentraxin domain characteristic of the family, and a unique N-terminal domain. It is produced by several cell types including endothelial and mononuclear cells. PTX3 acts as a pattern recognition receptor with non-redundant roles in the innate immune response to several microbial agents including the fungal pathogen Aspergillus fumigatus. Overexpression leads to enhanced pro-inflammatory responses. PTX3 is induced in response to LPS and inflammatory cytokines including TNF-alpha and IL-1 beta.

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.