Interleukin-1 (IL-1) is a family of proteins that includes classical members like IL-1α and IL-1β. IL- 1 receptor antagonist (IL-1RA), IL-18, IL-33, and IL-1F5-F10 share the same cell surface binding receptors as IL-1α and IL-1β and exhibit similar biological functions. IL-1β plays a crucial role in immune and inflammatory responses, bone remodeling, fever, carbohydrate metabolism, and the physiological processes of GH/IGF-1. Various diseases are associated with dysregulation or delayed expression of IL-1, including sepsis, rheumatoid arthritis, inflammatory bowel disease, acute and chronic myeloid leukemia, insulin-dependent diabetes, atherosclerosis, diseases related to nerve damage, and age-related degenerative diseases.

This assay kit employs the double antibody sandwich enzyme-linked immunosorbent assay (ELISA) technique. Specific anti-human IL-1β antibodies with high affinity are pre-coated onto the enzyme-linked plate. In the wells of the enzyme-linked plate, standards, test samples, and biotinylated detection antibodies are added. After incubation, IL-1β present in the samples binds to the solid-phase antibody and the detection antibody, forming an immune complex. After removing unbound substances, streptavidin-horseradish peroxidase labeled with horseradish peroxidase (HRP) is added. After wash step to remove any unbound substances, 3,3′,5,5′-Tetramethylbenzidine (TMB)substrate is added and color develops in proportion to the amount of IL-1β bound initially. The assay is stopped, and the optical density of the wells is determined using a micro-plate reader at 450nm wavelength (with a reference wavelength of 540nm or 570nm) for quantification.

Recovery
Human IL-1β of different levels was spiked into cell culture medium samples to determine its recovery rate. The recovery rate ranged from 85-107%, with an average recovery rate of 97%.
Sensitivity
The minimum detectable dose (MDD) of Human IL-1β is typically less than 1.0 pg/mL.

The MDD was determined by adding two standard deviations to the mean O.D. value of twenty zero standard replicates and calculating the corresponding concentration.
Linearity
To assess the linearity of the assay, four samples spiked with high concentrations of Human IL-1β were diluted with 1×Reagent Diluent to bring them within the detection range.

Dilution	Average % of Expected	Range (%)
1:2	102	98-107
1:4	103	101-107
1:8	106	103-110
1:16	107	105-113

Specificity
This assay recognizes natural and recombinant Human IL-1β.

The factors listed below were prepared at 50 ng/mL in 1×Reagent Diluent and assayed for cross-reactivity.

Preparations of the following factors prepared at 50 ng/mL in a mid-range Human IL-1β control were assayed for interference. No significant cross-reactivity or interference was observed.

Recombinant human protein	Recombinant mouse protein	Recombinant rat protein	Recombinant pig protein
IL-1α	IL-1α; IL-1β	IL-1α	IL-1α; IL-1β