Detects human IL-1 beta /IL-1F2 in Western blots. Shows less than 5% cross-reactivity with recombinant mouse (rm) IL‑1 beta and rpIL‑1 beta and no cross-reactivity with rrIL‑1 beta, rmIL‑1 alpha, rhIL‑1ra, rmIL‑1ra, or rrIL‑1 alpha.
Monoclonal Mouse IgG1 Clone # 8516
Protein A or G purified from hybridoma culture supernatant
E. coli-derived recombinant human IL-1 beta /IL-1F2 aa 117-269 Accession # P01584
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
<0.10 EU per 1 μg of the antibody by the LAL method.
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Human peripheral blood mononuclear cells treated with LPS, fixed with paraformaldehyde, and permeabilized with saponin
Measured by its ability to neutralize IL‑1 beta /IL‑1F2-induced proliferation in the D10.G4.1 mouse helper T cell line. Symons, J.A. et al. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 272. The Neutralization Dose (ND50) is typically 0.001-0.003 µg/mL in the presence of 50 pg/mL Recombinant Human IL‑1 beta /IL‑1F2 and 1.25 µg/mL concanavalin A.
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Cell Proliferation Induced by IL‑1 beta /IL‑1F2 and Neutralization by Human IL‑1 beta /IL‑1F2 Antibody.
Recombinant Human IL‑1 beta /IL‑1F2 (Catalog # 201-LB) stimulates proliferation in the the D10.G4.1 mouse helper T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL‑1 beta /IL‑1F2 (50 pg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti-Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB201). The ND50 is typically 0.001-0.003 µg/mL in the presence of concanavalin A (1.25 µg/mL).
Detection of Human IL‑1 beta /IL‑1F2 by Western Blot.
Western blot shows lysates of THP‑1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 24 hours and 10 µg/mL LPS and 3 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB201) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for IL‑1 beta /IL‑1F2 at approximately 36 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
IL‑1 beta /IL‑1F2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB201) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
IL‑1 beta /IL‑1F2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with LPS and monensin using Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB201) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (yellow; Catalog # NL007) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Preparation and Storage
Reconstitute at 0.5 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: IL-1 beta/IL-1F2
IL-1 is a name that designates two pleiotropic cytokines, IL-1 alpha (IL-1F1) and IL-1 beta (IL-1F2), which are the products of distinct genes. IL-1 alpha and IL-1 beta are structurally related polypeptides that share approximately 21% amino acid (aa) identity in human. Both proteins are produced by a wide variety of cells in response to inflammatory agents, infections, or microbial endotoxins. While IL-1 alpha and IL-1 beta are regulated independently, they bind to the same receptor and exert identical biological effects. IL-1 RI binds directly to IL-1 alpha or IL-1 beta and then associates with IL-1 R accessory protein (IL-1 R3/IL-1 R AcP) to form a high-affinity receptor complex that is competent for signal transduction. IL-1 RII has high affinity for IL-1 beta but functions as a decoy receptor and negative regulator of IL-1 beta activity. IL-1ra functions as a competitive antagonist by preventing IL-1 alpha and IL-1 beta from interacting with IL-1 RI (1 - 4). The human IL-1 beta cDNA encodes a 269 aa precursor. A 116 aa propeptide is cleaved intracellularly by the cysteine protease IL-1 beta -converting enzyme (Caspase-1/ICE) to generate the active cytokine (5-7). The 17 kDa mature human IL-1 beta shares 96% aa sequence identity with rhesus and 67-78% with canine, cotton rat, equine, feline, mouse, porcine, and rat IL-1 beta.
Allan, S.M. et al. (2005) Nat. Rev. Immunol. 5:629.
Boraschi, D. and A. Tagliabue (2006) Vitam. Horm. 74:229.
Kornman, K.S. (2006) Am. J. Clin. Nutr. 83:475S.
Isoda, K. and F. Ohsuzu (2006) J. Atheroscler. Thromb. 13:21.
March, C.J. et al. (1985) Nature 315:641.
Auron, P.E. et al. (1984) Proc. Natl. Acad. Sci. USA 81:7907.
Martinon, F. and J. Tschopp (2007) Cell Death Differ. 14:10.
R&D Systems personnel manually curate a database that contains references using R&D Systems products.
The data collected includes not only links to publications in PubMed,
but also provides information about sample types, species, and experimental conditions.
The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.