Human IL-1 beta /IL-1F2 Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-1 beta /IL-1F2 in sandwich ELISAs and Western blots. In sandwich ELISAs, less than 4% cross-reactivity with recombinant rat (rr) IL‑1 beta and less than 0.1% with recombinant porcine (rp) IL-1 beta, recombinant human IL-1 alpha, rpIL-1 alpha, rrIL-1 alpha, recombinant mouse (rm) IL-1 alpha, and rmIL-1 beta is observed.
  • Source
    Recombinant Monoclonal Mouse IgG1 Clone # 2805R
  • Purification
    Protein A or G purified from cell culture supernatant
  • Immunogen
    E. coli-derived recombinant human IL-1 beta /IL-1F2
  • Formulation
    Supplied as a solution in PBS. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Simple Western
    10 µg/mL
    See below
  • Immunocytochemistry
    8-25 µg/mL
    See below
    • Human IL-1 beta /IL-1F2 Sandwich Immunoassay
      Reagent
  • ELISA Capture (Matched Antibody Pair)
    2-8 µg/mL 
    Human IL‑1 beta /IL‑1F2 Antibody (Catalog # MAB601R)
  • ELISA Detection (Matched Antibody Pair)
    0.1-0.4 µg/mL 
    Human IL‑1 beta /IL‑1F2 Biotinylated Antibody (Catalog # BAF201)
  • ELISA Standard
     
    Recombinant Human IL-1 beta/IL-1F2 Protein (Catalog # 201-LB)
  • Neutralization
    Measured by its ability to neutralize IL‑1 beta /IL‑1F2-induced proliferation in the D10.G4.1 mouse helper T cell line. The Neutralization Dose (ND50) is typically 50-200 ng/mL in the presence of 50 pg/mL Recombinant Human IL‑1 beta /IL‑1F2.
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.
Data Examples
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 for 3 hours. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB601R) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for IL‑1 beta /IL‑1F2 at approximately 35 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
IL‑1 beta /IL‑1F2 in Human PBMCs.
IL‑1 beta /IL‑1F2 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) treated with 1 μg/mL LPS and
3 μM monensin for 24 hours using Mouse Anti-Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB601R) 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). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.
Detection of Human IL‑1 beta /IL‑1F2 by Simple WesternTM. Simple Western lane view shows lysates of THP‑1 human acute monocytic leukemia cell line untreated (-) or treated (+) with 200 nM PMA for 200 nM and 10 μg/mL LPS for 3 hours, loaded at 0.2 mg/mL. A specific band was detected for IL‑1 beta /IL‑1F2 at approximately 37 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human IL‑1 beta /IL‑1F2 Monoclonal Antibody (Catalog # MAB601R). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Cell Proliferation Induced by IL-1 beta /IL-1F2 and Neutralization by Human IL-1 beta /IL-1F2 Antibody. Recombinant HumanIL-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) as measured by Resazurin (Catalog # AR002). 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 # MAB601R). The ND50 is typically 50-200 ng/mL.
Preparation and Storage
  • Shipping
    The product is shipped with polar packs. 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 opening.
    • 6 months, -20 to -70 °C under sterile conditions after opening.
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.

  • References:
    1. Allan, S.M. et al. (2005) Nat. Rev. Immunol. 5:629.
    2. Boraschi, D. and A. Tagliabue (2006) Vitam. Horm. 74:229.
    3. Kornman, K.S. (2006) Am. J. Clin. Nutr. 83:475S.
    4. Isoda, K. and F. Ohsuzu (2006) J. Atheroscler. Thromb. 13:21.
    5. March, C.J. et al. (1985) Nature 315:641.
    6. Auron, P.E. et al. (1984) Proc. Natl. Acad. Sci. USA 81:7907.
    7. Martinon, F. and J. Tschopp (2007) Cell Death Differ. 14:10.
  • Long Name:
    Interleukin 1 beta
  • Entrez Gene IDs:
    3553 (Human); 16176 (Mouse); 24494 (Rat); 397122 (Porcine); 403974 (Canine); 100034237 (Equine); 100135556 (Guinea Pig)
  • Alternate Names:
    catabolin; IL1 beta; IL-1 beta; IL-1; IL1B; IL-1b; IL1-BETA; IL-1F2; IL1F2IL-1 beta; interleukin 1, beta; interleukin-1 beta; preinterleukin 1 beta; pro-interleukin-1-beta
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