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Česká republika Praha
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0032 (0)16 41 44 07
Recombinant Mouse Interleukin-1 Beta (IL-1β)
(Cat. No.: C042)
IL-1 is a name that designates two proteins, IL-1α and IL-1β, that are the products of distinct genes, but recognize the same cell surface receptors. IL-1α and IL-1β are structurally related polypeptides that show apprximately 25% homology at the amino acid level. Both proteins are produced by a wide variety of cells in response to stimuli such as those produced by inflammatory agents, infections, or microbial endotoxins. The proteins are synthesized as 31 kDa precursors that are subsequently cleaved into proteins with molecular weights of approximately 17.5 kDa. The specific protease responsible for the processing of IL-1β, designated interleukin 1β-converting enzyme (ICE), has been described. Mature human and mouse IL-1β share approximately 75% amino acid sequence identity and human
IL-1β has been found to be active on murine cell lines.
Recombinant Mouse IL-1 beta produced in E. coli is a non-glycosylated polypeptide chain containing 152 amino acids and having a molecular mass of 17400 Dalton.
Biological activity: The ED50 as determined by the dose-dependant stimulation of mouse D10S cells was found to be less than 0.01 ng/ml, corresponding to a Specific Activity of 1.0 x 108 IU/mg.
Purity:Greater than 98% 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 Val-Pro-Ile-Arg-Gln.
Endotoxin: Less than 0.1ng/µg (1 IEU/µg) of IL-1β.
Mouse IL-1β was lyophilized after extensive dialysis against PBS.
Lyophilized rmIL-1β although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution rmIL-1β should be stored at 4°C between 2-7 days and for future use below -18°C. For long-term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.
It is recommended to reconstitute the lyophilized rmIL-1β in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
1: Brain Behav Immun 2008 Dec;
Central inhibition of interleukin-1beta ameliorates sickness behavior in aged mice.
[Abstract] In elderly individuals high levels of interleukin-1beta (IL-1beta) in the brain have been implicated in infection-related behavioral pathologies but this has not been directly tested. Therefore, the current study investigated if sickness behavior in aged animals elicited by peripheral injection of lipopolysaccharide (LPS) is mediated through central IL-1beta. Adult and aged mice were injected intracerebroventricularly with either saline or IL-1ra (4mug) immediately prior to intraperitoneal administration of saline or LPS (10mug) and locomotor and social behaviors were assessed. As anticipated, LPS depressed locomotor activity and social behavior in both adult and aged mice but the behavioral deficits were markedly greater in the aged at 24h. Pretreatment with
IL-1ra did not affect LPS-induced sickness behavior in adults; however, in aged mice IL-1ra attenuated LPS-induced sickness behavior, restoring it to the level exhibited by young adults. Twenty-four hours post-injection hippocampal and hypothalamic tissues were collected to determine IL-1beta mRNA expression. Neither LPS nor IL-1ra affected IL-1beta mRNA levels in adults, presumably because any effect of LPS had dissipated by 24h. In contrast, IL-1beta mRNA was markedly higher in aged mice 24h after LPS, and prior treatment with IL-1ra either blocked or attenuated this effect in the hippocampus and hypothalamus, respectively. Taken together these data provide the first direct evidence that central IL-1beta is responsible for the severe sickness behavior observed in aged animals after LPS
treatment. Thus, inhibiting the central actions of IL-1beta may be useful for minimizing behavioral complications in older individuals with an infection.
2: Reprod Fertil Dev 2009 ;Vol 21(1)
108 effect of interleukin-1beta, -3 and -4 on bovine embryo development in vitro and MHC-I gene expression.
[Abstract] Studies in humans and other species have shown that the MHC class I region (MHC-I) is involved at a number of levels in the establishment and maintenance of pregnancy. Expression of MHC-I and MHC-I-like molecules is regulated during embryo development and differentiation. In general, classical MHC-I gene expression appears to be down-regulated in mammalian trophoblast cells, whereas the expression of certain non-classical (NC) MHC-I genes is up-regulated. Cytokines govern uterine immunology and receptivity and are increasingly recognized for their embryotrophic roles. The aim of the current study was to investigate the regulatory effect of a number of cytokines on bovine embryo development and NC MHC-I mRNA expression at the blastocyst stage. Cumulus oocyte
complexes were obtained by aspirating follicles from bovine ovaries collected from the local abattoir. At approximately 20 h postinsemination (hpi), presumptive zygotes produced following IVM/IVF were denuded by gentle vortexing and transferred to synthetic oviduct fluid medium supplemented with different concentrations (0, 0.1, 1, 10, 100 ng mL(-1)) of either Interleukin (IL)-1beta (n = 1759), IL-3 (n = 1053) or IL-4 (n = 1549) and cultured in an atmosphere of 5% CO(2), 5% O(2), and 90% N(2) at 100% humidity. Cleavage rates were recorded at 48 hpi and the proportion of embryos developing to the blastocyst stage was recorded from Day 6 to 8. Day 7 blastocysts were snap frozen in pools of 10 and stored at -80 degrees C for the analysis of transcript abundance of bovine NC MHC-I genes
(BOLA-NC1, NC2, NC3 and NC4, see www.ebi.ac.uk/ipd/mhc/) using quantitative real-time RT-PCR. Messenger RNA was extracted from 5 pools of 10 embryos per treatment, using Dynabeads mRNA DIRECTMicro Kit (Dynal A.S, Oslo, Norway), and cDNA was synthesized. Real-time PCR was used to compare NC MHC-I transcript abundance. The data were analyzed using the standard curve method, and means were compared by the Student t-test. There was no effect of any of the cytokines tested on cleavage or blastocyst development. However, the relative abundance of the BOLA-NC1 transcript significantly increased (P </= 0.05) 1.6-fold and 3-fold, respectively, as the concentration of IL-3 and IL-4 in culture increased, whereas there was no significant change (P >/= 0.05) in the abundance of IL-1beta. There was no
significant difference (P >/= 0.05) in the levels of BOLA NC2 and BOLA-NC4 in the embryos cultured with IL-3 and IL-1beta, irrespective of the concentration, but the levels varied significantly (P </= 0.05) at higher (>/=10 ng mL(-1)) concentrations of IL-4, 2-fold (NC2) and 3-fold (NC4). The relative abundance of the BOLA-NC3 transcript significantly increased (1.5-fold, P </= 0.05) as the concentration of IL-3 in culture increased, whereas there was no significant change (P >/= 0.05) in the presence of IL-1beta or IL-4. Compared with the lack of differences in embryo developmental competence, the NC MHC-I expression data possibly suggest a preferential immunomodulatory role of these cytokines during preimplantation embryo development.
3: Vet Ophthalmol ;Vol 12(1)
Anti-inflammatory effects of topical supernatant from human amniotic membrane cell culture on canine deep corneal ulcer after human amniotic membrane transplantation.
[Abstract] The objective of this study was to examine the effect of topically applied human amniotic epithelial cell (HAEC) culture supernatant on corneal inflammatory reaction in dogs. Twenty-five dogs were randomly assigned into five groups. The control group consisted of five dogs with normal cornea. Inductions of corneal ulcers were performed using 0.45 cm trephine and human amniotic membrane was transplanted in 20 dogs. These 20 dogs were assigned into four treatment groups: topical antibiotic, topical corticosteroid, topical mock media and topical culture supernatant from HAEC, respectively. Administrations of the testing agents started at 24 h (h) after transplantation four times daily for nine consecutive days. Tears were collected before an operation 24 h after
transplantation, but before application of the testing agents on consecutive odd days following transplantation. The concentrations of interleukin-1beta (IL-1beta) and nitric oxide (NO) in tear fluid were measured using canine IL-1beta ELISA kit and Griess assay, respectively. Our analysis indicates that elevations of IL-1beta and NO concentrations are associated with inflammatory conditions in the eyes. Corticosteroid, a reference anti-inflammatory drug, and the culture supernatant from HAEC significantly decreased IL-1beta and NO concentrations. In addition, the clinical signs such as conjunctivitis and neovascularization were decreased in both topical corticosteroid and supernatant from HAEC treated groups. Mock and antibiotic solutions failed to decrease NO and IL-1beta concentrations.
In conclusion, topical application of the culture supernatant from HAEC alleviated inflammation in induced-corneal ulcer of dogs, possibly via inhibition of IL-1beta and NO production.
4: 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.
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