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.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
The recovery of sgp130 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=5)
Citrate Plasma (n=5)
EDTA Plasma (n=5)
Heparin Plasma (n=5)
To assess the linearity of the assay, five samples were spiked with high concentrations of sgp130 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
Refer to the product for 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.
Prepare all reagents, standard dilutions, and samples as directed in the product insert.
Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 3 hours.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
Add 200 µL of Conjugate to each well.
For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour. For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
Aspirate and wash 4 times.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
50 µL Stop Solution
Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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