Human Soluble gp130 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human soluble gp130 Immunoassay is a 4.5 or 5.5 hour solid phase ELISA designed to measure sgp130 in cell culture supernates, serum, and plasma. It contains soluble insect cell-expressed recombinant human gp130 and antibodies raised against recombinant human gp130 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural gp130 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 gp130.

Preparation and Storage

Background: gp130

Gp130, the common signal transducing receptor component shared by the functional receptor complexes of the IL-6 family of cytokines, belongs to the classI cytokine receptor family. Binding of IL-6 or IL-11 to either the membrane-anchored or soluble IL-6 R or IL-11 R, respectively, initiates the association of IL-6 R (IL-11 R) with gp130. The complex then undergoes homodimerization and signal transduction. For other IL-6 family cytokines, such as LIF and OSM, signal transduction is similarly triggered by the heterodimerization of gp130 with LIFR or OSM R.

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.