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Recombinant Human Interleukin-1 Receptor Antagonist (IL-1RN)
(Cat. No.: C033)
Background:
Interleukin-1 receptor antagonist (IL-1RN) is a member of the IL-1 family. Endogenous IL-1RN is produced in numerous animal disease models as well as in human autoimmune and chronic inflammatory diseases. It binds to IL-1 receptors in competition with IL-1, but does not elicit intracellular response from this binding. Its role in counteracting the proinflammatory effects of IL-1 is being studied by numerous research groups.
IL-4 and IL-13 have been shown to amplify the stimulatory effect of IL1-beta on the production of soluble and intracellular forms of IL-1RN.
The regulated expression of IL-1RN in various cell types has been shown to be influenced by cytokines. In synovial fibroblasts the synthesis of IL-1RN is markedly enhanced by IL-1, TNF-alpha, or PDGF.
Description:
Recombinant Human IL-1RN produced in E. coli is a non-glycosylated, N-terminal methionyl form of the human naturally-occurring polypeptide chain containing 153 amino acids and having a molecular mass of 17258 Dalton.
Quality Control:
Biological activity: The ED50 as determined by the dose-dependant inhibition of IL-1 stimulation of D10S cells was found to be 0.5 ng/ml, corresponding to a Specific Activity of 2.0 x 106 IU/mg.
Purity: Greater than 95% as determined by
(a) Analysis by RP-HPLC.
(b) Anion-exchange FPLC.
(c) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel..
Amino-Acid Sequence: The sequence of the first five N-terminal amino acids was determined and was found to be Met-Arg-Pro-Ser-Gly.
Endotoxin: Less than 0.1ng/µg (1IEU/µg) of IL-1RN.
Formulation: IL-1RN was lyophilized after extensive dialysis against PBS.
Storage: Lyophilized rHuIL-1RN although stable at room temperature for 3 weeks, should be stored desiccated below -18oC. Upon reconstitution rHuIL-1RN should be stored at 4oC between 2-7 days and for future use below -18oC. For long-term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.
Reconstitution: It is recommended to reconstitute the lyophilized rHuIL-1RN in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Latest Publications:
1: Rheumatol Int 2009 Jan;
Genetic polymorphisms of interleukin-1beta (-511C/T) and interleukin-1 receptor antagonist (86-bpVNTR) in susceptibility to knee osteoarthritis in a Chinese Han population.
[Abstract] Osteoarthritis (OA) is a multifactorial disorder in which genetic factors act as important contributors to its onset and progression. Associations between genetic polymorphisms of the interleukin-1 (IL-1) gene cluster and OA susceptibility have been studied continuously in different ethnic groups, yielding controversial results. This study investigated the association of interleukin-1beta (-511C/T) and interleukin-1 receptor antagonist (86-bp VNTR) polymorphisms with knee OA susceptibility in a Chinese Han population. A case-control association study was conducted. The two polymorphisms were genotyped in 453 patients who had primary symptomatic knee OA with radiographic confirmation and in 487 matched controls. Allelic and genotypic frequencies and haplotype
distribution were compared between OA and control subjects. For either of the two loci, no significant difference was detected in genotype or allele distribution between knee OA and control groups (all P > 0.05). The haplotype distribution of the two loci showed no difference between the two groups, either. Furthermore, no association between the genotype of the -511 and VNTR polymorphisms and the clinical variables, age, sex, body mass index and Kellgren/Lawrence score was observed in OA patients. The genetic polymorphisms of interleukin-1beta and interleukin-1 receptor antagonist are not risk factors for OA etiology in Han Chinese.
2: Arq Bras Cardiol 2008
Nov;Vol 91(5)
Interleukin-1 receptor antagonist gene VNTR polymorphism is associated with coronary artery disease.
[Abstract] BACKGROUND: Coronary Artery Disease (CAD) is the atherosclerosis of coronary arteries that carry blood to the heart muscle. Atherosclerosis is an inflammatory disease. Cytokine gene variations such as those associated with the IL1 family are involved in the pathogenesis of atherosclerosis. OBJECTIVE: The purpose of this study was to determine the relationship between IL1 family polymorphisms (IL1RN VNTR, IL1B positions -511 and +3953) and CAD in Turkish population. METHODS: 427 individuals were submitted to coronary angiography and were grouped as 170 control subjects and 257 CAD patients. The CAD subjects were divided into two subgroups: 91 Single Vessel Disease (SVD) and 166 Multiple Vessel Disease (MVD) subjects. The genotypes of IL1RN and of IL1B (-511,
+3953) were determined by polymerase chain reaction (PCR) followed by restriction digestion analysis. RESULTS: No significant difference was found in IL1RN and IL1B (-511 and +3953) genotype distributions between CAD and control subjects or MVD and control subjects. However, significant association was seen in IL1RN 2/2 genotype between SVD and control subjects (P= 0.016, x2: 10.289, OR: 2.94, 95% CI: 1.183-7.229). Similarly, no statistically significant difference was found in IL1RN and IL1B (-511 and +3953) allele frequencies between CAD and control subjects, MVD and control subjects or SVD and control subjects. CONCLUSION: No association was found in either allele frequency or genotype distribution of IL1RN and IL1B polymorphisms between CAD and the control groups. However; IL1RN 2/2
genotype may be a risk factor for SVD in the Turkish population.
3: Pediatr Neurol 2009 Feb;Vol 40(2)
Febrile Seizures: Interleukin 1beta and Interleukin-1 Receptor Antagonist Polymorphisms.
[Abstract] In order to investigate the association between IL-1beta -511 C-->T and IL-1 receptor antagonist intron 2 variable tandem repeat polymorphisms, and febrile seizures in children, 90 children (mean age, 19.7 +/- 11.2 months) diagnosed with febrile seizure and 106 healthy controls (mean age, 14.2 +/- 3.6 months) with no seizure or neurologic events were included in the study. The polymorphisms were analyzed using restriction fragment length polymorphism and agarose gel electrophoresis methods. In the patient group, the frequencies of IL-1beta genotypes CC, CT, and TT were 24.4%, 52.2%, and 23.3%, respectively, compared with 38.7%, 50.95%, and 10.4%, respectively, in the control group. The TT genotype was significantly more common in the patient group than in the
control group (P = 0.044), and the T allele frequency was significantly higher in the patient group (0.50 vs 0.36, P = 0.040). Among the three genotypes (RN1/1, RN1/2, and RN2/2) of the IL receptor antagonist gene variable tandem repeat polymorphisms, the frequency of both the RN2/2 genotype and the RN2 allele were significantly higher in the patient group (P = 0.007). Also RN2 allele frequency was found higher in patient group than controls (0.29 vs 0.15, P = 0.020). IL-1beta -511 and IL-1 receptor antagonist intron 2 variable tandem repeat polymorphisms may be involved in susceptibility to febrile convulsions in children.
4: Biophys J 2009 Jan;Vol 96(1)
Multistep aggregation pathway of human interleukin-1 receptor antagonist: kinetic, structural, and morphological characterization.
[Abstract] The complex, multistep aggregation kinetic and structural behavior of human recombinant interleukin-1 receptor antagonist (IL-1ra) was revealed and characterized by spectral probes and techniques. At a certain range of protein concentration (12-27 mg/mL) and temperature (44-48 degrees C), two sequential aggregation kinetic transitions emerge, where the second transition is preceded by a lag phase and is associated with the main portion of the aggregated protein. Each kinetic transition is linked to a different type of aggregate population, referred to as type I and type II. The aggregate populations, isolated at a series of time points and analyzed by Fourier-transform infrared spectroscopy, show consecutive protein structural changes, from intramolecular
(type I) to intermolecular (type II) beta-sheet formation. The early type I protein spectral change resembles that seen for IL-1ra in the crystalline state. Moreover, Fourier-transform infrared data demonstrate that type I protein assembly alone can undergo a structural rearrangement and, consequently, convert to the type II aggregate. The aggregated protein structural changes are accompanied by the aggregate morphological changes, leading to a well-defined population of interacting spheres, as detected by scanning electron microscopy. A nucleation-driven IL-1ra aggregation pathway is proposed, and assumes two major activation energy barriers, where the second barrier is associated with the type I --> type II aggregate structural rearrangement that, in turn, serves as a pseudonucleus
triggering the second kinetic event.
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