Human IL-1RA/IL-1F3 DTSet enzyme-linked immunoassay kit
| Specification | 96*5 Test；96T*15 Test |
|---|---|
| Standard Curve Range | 31.25 pg/ml -2000 pg/ml |
| Standard Curve Gradient | 7 Points/3 Folds |
| Number of Incubations | 2 |
| Detectable sample | |
| Sample Volume | 50 μl |
| Type | Not Ready-to-Use |
| Test Duration | 120min |
| pg/ml | O.D. | Average | Corrected | |
|---|---|---|---|---|
| 0.00 | 0.0704 | 0.0690 | 0.0697 | |
| 31.25 | 0.3218 | 0.2896 | 0.3057 | 0.2360 |
| 62.50 | 0.4407 | 0.5047 | 0.4727 | 0.4030 |
| 125.00 | 0.7050 | 0.7112 | 0.7081 | 0.6384 |
| 250.00 | 1.2660 | 1.2780 | 1.2720 | 1.2023 |
| 500.00 | 2.2600 | 2.1290 | 2.1945 | 2.1248 |
| 1000.00 | 3.3460 | 2.9920 | 3.1690 | 3.0993 |
| 2000.00 | 4.0657 | 4.0810 | 4.0734 | 4.0037 |
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Economical alternative to complete kits
Kit Content
- Capture Antibody
- Detection Antibody
- Recombinant Standard
- Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)
Other Reagents Required
DTSet Ancillary Reagent Kit (5 plates): containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and assay buffer.
- 96 well microplates: YOUKE Life, Catalog # DSEP01. Plate Sealers: YOUKE Life, Catalog # DSSF01.
- Coating Buffer: 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2-7.4, 0.2μm filtered . YOUKE Life, Catalog # DSCB01.
- Blocking Buffer: YOUKE Life, Catalog # DSBB01.
- Wash Buffer: 0.05% Tween® 20 in PBS, pH 7.2-7.4. YOUKE Life, Catalog # DSWB01.
- Assay Buffer: 0.5% BSA，0.05% Tween® 20，PBS Solution.YOUKE Life, Catalog # DSAB01
- Substrate Solution: Tetramethylbenzidine. YOUKE Life, Catalog # DSTS01.
- Stop Solution: 0.5mol/ml H2SO4. YOUKE Life, Catalog # DSSS01.
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.
