Human IL-1RA/IL-1F3 enzyme-linked immunoassay kit
| Specification | 96 Test |
|---|---|
| Sensitivity | 0.09 pg/ml (50 μl)；0.15 pg/ml (10 μl)； |
| Standard Curve Range | 2.74~2000 pg/ml |
| Standard Curve Gradient | 7 Points/3 Folds |
| Number of Incubations | 2 |
| Detectable sample | Liquid phase sample of soluble substances. For example: serum, plasma, cell culture supernatant, tissue grinding liquid, etc. |
| Sample Volume | 50 μl/10 μl |
| Type | Ready-to-Use |
| Operation Duration | 120min |
| pg/ml | O.D. | Average | Corrected | |
|---|---|---|---|---|
| 0.00 | 0.0625 | 0.0611 | 0.0618 | |
| 2.74 | 0.0946 | 0.1014 | 0.0980 | 0.0362 |
| 8.23 | 0.1404 | 0.1366 | 0.1385 | 0.0767 |
| 24.69 | 0.2081 | 0.1979 | 0.2030 | 0.1412 |
| 74.07 | 0.3790 | 0.3737 | 0.3764 | 0.3146 |
| 222.22 | 0.9861 | 0.9935 | 0.9898 | 0.9280 |
| 666.67 | 2.1300 | 2.1710 | 2.1505 | 2.0887 |
| 2000.00 | 3.6470 | 3.7770 | 3.7120 | 3.6502 |
Precision
| Intra-assay Precision | Inter-assay Precision | |||||
| Sample Number | S1 | S2 | S3 | S1 | S2 | S3 |
| 22 | 22 | 22 | 6 | 6 | 6 | |
| Average(pg/ml) | 37.9 | 190.5 | 584.1 | 42.1 | 208.8 | 611.7 |
| Standard Deviation | 2.8 | 12.1 | 37.9 | 1.9 | 14.5 | 41.6 |
| Coefficient of Variation(%) | 7.4 | 6.3 | 6.5 | 5.3 | 4.2 | 4.6 |
Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.
Spike Recovery
The spike recovery was evaluated by spiking 3 levels of human IL-1RA into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 85% to 109% with an overall mean recovery of 98%.
Sample Values
| Sample Matrix | Sample Evaluated | Range (ng/ml) | Detectable (%) | Mean of Detectable (ng/ml) |
|---|---|---|---|---|
| Serum | 30 | 6.34-126.18 | 100 | 33.78 |
Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of IL-1RA in this assay. No medical histories were available for the donors.
Product Data Sheet
Background: IL-1RA/IL-1F3
Interleukin-1 receptor antagonist (IL-1ra; also known as IL-1F3) is a 22-25 kDa member of the IL-1 family of cytokines. Currently, there are 11 family members (IL-1F1-F11), nine of which form an IL-1 gene cluster on human Ch2. Each IL-1 family member contains an IL-1 fold. This fold is generated by 12 packed beta -sheets that interact to form a beta -trefoil structure. Little amino acid (aa) homology is required to achieve this structure, and this explains the low aa identity among IL-1 family members. IL-1ra is a pure cytokine receptor antagonist that has no signal transduction-initiating activity. It is an acute phase protein that exists to dampen inflammation. IL-1( beta ) is initially produced by monocytes in response to a variety of stimuli. Circulating IL-1 then binds to widely expressed IL-1 type I receptors (IL-1 RI) and initiates a number of pro-inflammatory events. On endothelial cells (EC), IL-1 induces PGE2 and IL-6 release, generating fever, thrombocytosis, and hepatic acute phase protein production. In synovial joints, IL-1 induces chondrocyte NO production, an event that leads to reduced collagen synthesis and chondrocyte apoptosis. Finally, IL-1 increases neutrophil counts, both in blood and tissue, and thus is able to promote a pro-inflammatory environment in multiple locations. IL-1ra blocks IL-1 action through competitive inhibition. More correctly, although IL-1ra fills the IL-1 binding site in IL-1 RI, it is also unable to orchestrate the creation of a signal-transducing IL-1 RI:IL-1 R Accessory protein (IL-1 R AcP) heterodimer complex. Effective IL-1ra concentrations are generally 100-fold greater than local IL-1 concentrations. This is because the IL-1ra half-life is but 6 minutes, and very few IL-1 type I receptors need to be engaged by IL-1 to elicit a cellular response.
Human IL-1ra is synthesized as a 177 aa precursor that contains a 25 aa signal sequence and a 152 aa mature region. Although it contains an IL-1 cytokine fold, it apparently lacks two structural motifs that allow for activation of the IL-1 receptor heterodimer. First, and following binding to IL-1 RI, the presence of Ile 51-His 54 and Lys 145 of the mature molecule preclude recruitment of IL-1 R AcP. Second, there is no identifiable C-terminal lectin segment that is hypothesized to help recruit an accessory signaling component. Mature human IL-1ra is 77% and 82% aa identical to mouse and canine IL-1ra, respectively, and human IL-1ra inhibits IL-1 activity on mouse cells. A number of cell types express IL-1ra, including monocytes, Sertoli cells, hepatocytes, adipocytes, synovial fibroblasts, mast cells, pancreatic beta -cells, and intestinal epithelial cells. There are at least three intracellular IL-1ra isoforms (icIL-1ra1, 2, and 3). All show N-terminal variation, and all contain amino acids 35-177 of the secreted precursor. Intracellular IL-1ra1 is of particular interest, because it is reported to be "secreted" by endothelial cells and binds to the IL-1 RI in an antagonist fashion. Intracellular IL-1ra1 is 159 aa in length and shows a 3 aa substitution for the first 21 aa's of the signal sequence of IL-1ra.
