CellRapeutics™ Human IL-6 ELISA Kit
The Human IL-6 Immunoassay is a 4.5 hour solid phase ELISA designed to measure IL-6 in cell culture supernates, serum, and plasma. It contains recombinant human IL-6 and antibodies raised against recombinant human IL-6 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural IL-6 showed linear curves that were parallel to the standard curves obtained using the E. coli-expressed Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human IL-6.
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.
Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma
Cell Culture Supernates
The recovery of human IL-6 spiked to three different levels throughout the range of the assay in various matrices was evaluated.
To assess the linearity of the assay, four samples were spiked with high concentrations of IL-6 in various matrices and diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
IL-6 (Interleukin-6) is a pleiotropic cytokine that acts in the acute phase reaction, inflammation, hematopoiesis, bone metabolism, and cancer progression. It contributes to chronic inflammation in obesity, insulin resistance, inflammatory bowel disease, arthritis, sepsis, and atherosclerosis. IL-6 signals through a receptor complex of IL-6 R alpha and gp130. gp130 is also a component of the receptors for CLC, CNTF, CT-1, IL-11, IL-27, LIF, and OSM. Soluble forms of IL-6 R alpha are generated by both alternative splicing and proteolytic cleavage. In a mechanism known as trans-signaling, complexes of soluble IL-6 and IL-6 R alpha elicit responses from gp130-expressing cells that lack cell surface IL-6 R alpha. Long Name Interleukin 6 Entrez Gene IDs 3569 (Human); 16193 (Mouse); 24498 (Rat); 399500 (Porcine); 280826 (Bovine); 403985 (Canine); 100034196 (Equine); 493687 (Feline); 100008733 (Rabbit); Alternate Names B cell stimulatory factor-2; B-cell differentiation factor; BSF-2; BSF2CTL differentiation factor; CDF; HGFHSFIFNB2Hybridoma growth factor; IFN-beta 2; IFN-beta-2; IL-6B-cell stimulatory factor 2; Interferon beta-2; interleukin 6 (interferon, beta 2); interleukin BSF-2; interleukin-6; MGI-2A;
Product format/Physical form
96-well strip plate
Solid Phase Sandwich ELISA
3.1 - 300 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
Natural and recombinant human IL-6
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
No significant interference observed with available related molecules.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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.
100 µL Assay Diluent
3. Add 100 µL of Assay Diluent to each well.
100 µL Standard, Control, or Sample
4. Add 100 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 3 times for a total of 4 washes.
200 µL Conjugate
6. Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.
7. Aspirate and wash 4 times.
200 µL Substrate Solution
8. Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
50 µL Stop Solution
9. 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.
For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.