Porcine IL-1 beta/IL-1F2 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Porcine IL-1 beta Immunoassay is a 4.5 hour solid phase ELISA designed to measure porcine IL-1 beta in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant mature porcine IL-1 beta and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant porcine IL-1 beta. Results obtained using natural porcine IL-1 beta show dose response curves that are 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 porcine IL-1 beta.

Preparation and Storage

Background: IL-1 beta/IL-1F2

IL-1 beta (Interleukin-1 beta) is produced in response to inflammatory agents, infections, or microbial endotoxins. It plays a central role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and GH/IGF-I physiology. IL-1 beta dysregulation is implicated in many pathological conditions including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myelogenous leukemia, insulin-dependent diabetes mellitus, atherosclerosis, neuronal injury, and aging-related diseases. Intracellular cleavage of the IL-1 beta precursor by Caspase-1/ICE is a key step in the inflammatory response. IL-1 beta binds to IL-1 RI and IL-1 RII. The IL-1 receptor accessory protein (IL-1 RAcP) associates with IL-1 RI and is required for IL-1 RI signal transduction, and IL-1ra is a secreted molecule that functions as a competitive inhibitor of IL-1 bioactivity.

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.