Human IL-36 alpha /IL-1F6 Antibody

Recombinant Monoclonal Antibody.
  
  • Species Reactivity
    Human
  • Specificity
    Detects human IL-36 alpha /IL-1F6 in direct ELISAs and Western blots.
  • Source
    Recombinant Monoclonal Rabbit IgG Clone # 1087L
  • Purification
    Protein A or G purified from hybridoma culture supernatant
  • Immunogen
    E. coli-derived recombinant human IL-36 alpha /IL-1F6
    Lys6-Phe158
    Accession # Q9UHA7
  • Formulation
    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.
  • 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
  • Neutralization
    Measured by its ability to neutralize IL‑36 alpha /IL‑1F6-induced IL-8 secretion in the A431 human epithelial carcinoma cell line. The Neutralization Dose (ND50) is typically 0.2-1.2 µg/mL in the presence of 80 ng/mL Recombinant Human IL‑36 alpha /IL‑1F6 (aa 6-158).
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‑36 alpha /IL‑1F6 by Western Blot. Western blot shows lysates of HEK293T human embryonic kidney cell line transfected with human IL-36 alpha /IL-1F6. PVDF membrane was probed with 1 µg/mL of Rabbit Anti-Human IL‑36 alpha /IL‑1F6 Monoclonal Antibody (Catalog # MAB10781) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for IL‑36 alpha /IL‑1F6 at approximately 15 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Il-8 Secretion Induced by IL‑36 alpha /IL‑1F6 and Neutralization by Human IL‑36 alpha /IL‑1F6 Antibody. Recombinant Human IL‑36 alpha /IL‑1F6 (aa 6-158) stimulates CXCL8/IL-8 secretion in the A431 human epithelial carcinoma cell line in a dose-dependent manner (orange line), as measured by the Human CXCL8/IL-8 Quantikine ELISA Kit (Catalog # D8000C).CXCL8/
IL-8 secretion elicited by Recombinant Human IL‑36 alpha /
IL‑1F6 (aa 6-158) (80 ng/mL) is neutralized (green line) by increasing concentrations of Rabbit Anti-Human IL‑36 alpha /IL‑1F6 Monoclonal Antibody (Catalog # MAB10781). The ND50 is typically 0.2-1.2 µg/mL.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.5 mg/mL in sterile PBS.
  • Shipping
    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-36 alpha/IL-1F6

Human IL-36 alpha, previously called IL-1F6 and FIL1 epsilon  (family of IL-1 member epsilon), is a member of the IL-1 family which includes IL-1 beta, IL-1 alpha, IL-1ra, IL-18, and novel family members IL-36 Ra (IL-1F5), IL-36 beta (IL-1F8), IL-36 gamma (IL-1F9), IL-37 (IL-1F7) and IL‑38 (IL‑1F10) (1‑4). All family members show a 12  beta ‑strand, beta -trefoil configuration, and are believed to have arisen from a common ancestral gene (1, 2). IL-36 alpha is an 18‑22 kDa, 158 amino acid (aa) intracellular and secreted protein that contains no signal sequence, no prosegment and no potential from N‑linked glycosylation sites (1‑3). It can be released in response to LPS and the cell ATP‑induced activation of the P2X7 receptor (5). A 120 aa isoform missing aa 1‑38 has been reported (6). Human IL‑36 alpha (aa 6 ‑ 158) shares 57‑68% aa sequence identity with mouse, rabbit, equine and bovine IL‑36 alpha and 27‑57% aa sequence identity with other novel IL‑1 family members. IL‑36 alpha is mainly found in skin and lymphoid tissues, but also in fetal brain, trachea, stomach and intestine (1, 3, 7). It is expressed by monocytes, B and T cells (1, 2). The receptor for IL‑36 alpha is a combination of IL‑1 Rrp2 (also called IL1RL2 or IL‑1 R6), mainly found in epithelia and keratinocytes, and the widely expressed IL‑1 RAcP (3, 7). IL-36 alpha, beta, and gamma all activate NF-kappa B and MAPK pathways in an IL‑1 Rrp2 dependent manner, and induce production of inflammatory cytokines and chemokines such as CXCL8/IL-8 (7). IL-36 alpha and other family members are overexpressed in psoriatic skin lesions, and transgenic overexpression of IL‑36 alpha in skin keratinocytes produces epidermal hyperplasia (7‑9). IL-36 alpha is present in kidney tubule epithelia, and it is highly expressed in intubulointerstitial lesions in mouse models of chronic glomerulonephritis, lupus nephritis and diabetic nephritis (10).

  • References:
    1. Smith, D.E. et al. (2000) J. Biol. Chem. 275:1169.
    2. Dunn, E. et al. (2001) Trends Immunol. 22:533.
    3. Barksby, H.E. et al. (2007) Clin. Exp. Immunol. 149:217.
    4. Dinarello, C. et al. (2010) Nat. Immunol. 11:973.
    5. Martin, U. et al. (2009) J. Immunol. 183:4021.
    6. Entrez Accession # EAW73614.
    7. Towne, J.E. et al. (2004) J. Biol. Chem. 279:13677.
    8. Blumberg, H. et al. (2010) J. Immunol. 185:4354.
    9. Johnston, A. et al. (2011) J. Immunol. 186:2613.
    10. Ichii, O. et al. (2010) Lab. Invest. 90:459.
  • Long Name:
    Interleukin 36 alpha/Interleukin 1 Family 6
  • Entrez Gene IDs:
    27179 (Human); 54448 (Mouse)
  • Alternate Names:
    FIL1 epsilon; FIL1; FIL1(EPSILON); FIL1E; IL-1 epsilon; IL1(EPSILON); IL1E; IL-1E; IL-1F6 (FIL-1-epsilon); IL1F6; IL-1F6; IL36 alpha; IL-36 alpha; IL36A; interleukin 1 family, member 6 (epsilon); interleukin 1, epsilon; Interleukin 36, Alpha; Interleukin-1 epsilon; interleukin-1 family member 6; Interleukin-36 Alpha
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