Human B7-H3 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human B7-H3 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human B7-H3 in cell culture supernates, serum, plasma, and urine. It contains NS0-expressed recombinant human B7-H3 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human B7-H3 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 naturally occurring human B7-H3.

Preparation and Storage

Background: B7-H3

B7-H3 is an immunoglobulin superfamily protein that is expressed by osteoblasts, fibroblasts, ciliated respiratory epithelial cells, anterior pituitary progenitor cells, first trimester extravillous trophoblasts, and multiple tumor cells. It is induced on activated dendritic cells (DC), skeletal muscle myocytes, and monocytes. B7-H3 can both stimulate and inhibit immune cell activation. As a co-stimulator, B7-H3 augments macrophage activation and promotes proliferation and cytokine secretion by CD4+ and CD8+ T cells. As a co-inhibitor, tumor cell B7-H3 blocks the cytolytic activity of NK cells, limits cytokine production by activated T cells, and imparts an immunosuppressive phenotype to dendritic cells. B7-H3 can interact with TREML2/TLT-2 to enhance T cell activation including the CD8+ T cell-mediated lysis of tumor cells.

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.