Rat IL-9 Antibody

  
  • Species Reactivity
    Rat
  • Specificity
    Detects rat IL-9 in direct ELISAs and Western blots. In direct ELISAs, approximately 30% cross-reactivity with recombinant mouse IL-9 is observed, and approximately 10% cross-reactivity with recombinant human IL-9 is observed.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant rat IL‑9
    Gln19-Ala144
    Accession # NP_001099217
  • 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
    0.1 µg/mL
    Recombinant Rat IL‑9 (Catalog # 4134-RL)
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
  • Neutralization
    Measured by its ability to neutralize IL‑9-induced proliferation in the TS1 mouse helper T cell line. Moreau, F. J. et al. (1988) Nature 336:690. The Neutralization Dose (ND50) is typically 1.5-7.5 µg/mL in the presence of 10 ng/mL Recombinant Rat IL‑9.
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
Cell Proliferation Induced by IL‑9 and Neutralization by Rat IL‑9 Antibody. Recombinant Rat IL‑9 (Catalog # 4134-RL) stim­ulates proliferation in the TS1 mouse helper T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL‑9 (10 ng/mL) is neutralized (green line) by increasing concentrations of Sheep Anti-Rat IL‑9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4134). The ND50 is typically 1.5‑7.5 µg/mL.
Intracellular Staining by Flow Cytometry
Detection of IL‑9 in Rat Spleno­cytes by Flow Cytometry. Rat splenocytes were treated for 24 hours with 50 ng/mL PMA and 500 ng/mL Ca2+ ionomycin then stained with Sheep Anti-Rat IL‑9 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4134, filled histogram) or control antibody (Catalog # 5‑001‑A, open histogram), followed by NorthernLights™ 637-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # NL011). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permea­bilized with saponin.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 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-9

Rat interleukin-9 (IL-9; also T-cell growth factor P40 and MEA) is a 20‑30 kDa secreted glycoprotein that belongs to the IL-7/IL-9 family of hematopoietic cytokines. It is best considered an atypical Th2-type cytokine (1‑4). Rat IL-9 was initially isolated from the 4437A rat thymoma cell line (5). The IL-9 precursor is 144 amino acids (aa) in length and contains an 18 aa signal sequence plus a 126 aa mature segment (5). An extended, 219 aa alternate precursor form is reported in GenBank (Accession # XP_001068265). It shows a 113 aa substitution for the first 38 aa of the standard form. Its significance is unknown. Mature rat IL-9 is 75%, 74% and 58% aa identical to cotton rat, mouse and human IL-9, respectively (6‑9). There is variability in cross-species reactivity. Canine and mouse IL-9 are purportedly active on human cells (9‑11), while human IL-9 is inactive on mouse cells (9). Cells reported to express IL-9 include activated CD4+ T cells and eosinophils (12, 13). The receptor for rat IL-9 is composed of the 62 kDa rat IL-9R alpha chain and the 64 kDa rat common cytokine receptor gamma -chain (5, 14, 15). IL-9 is suggested to play a significant, but likely supporting role in allergy and asthma. Its synthesis is induced upon exposure to allergen (10, 13). Following release, it is known to upregulate IgE production by IL‑4‑stimulated B cells (16). It also stimulates mucin secretion and IL-13 production through a direct action on respiratory epithelium (10, 17). In the bone marrow, it promotes eosinophilia, likely through IL-5 induction (18, 19). And once produced, eosinophils are chemoattracted to the bronchi through the IL-9-induced secretion of eotaxin-1 by airway smooth muscle cells (20).

  • References:
    1. Renauld, J-C. et al. (1993) Adv. Immunol. 54:79.
    2. Demoulin J-B. and J-C. Renauld (1998) Intern. Rev. Immunol. 16:345.
    3. Knoops, L. and J-C. Renauld (2004) Growth Factors 22:207.
    4. Finiasz, M.R. et al. (2006) Clin. Exp. Immunol. 147:139.
    5. Flubacher, M.M. et al. (1994) J. Virol. 68:7709.
    6. Blanco, J.C. et al. (2004) J. Interferon Cytokine Res. 24:21.
    7. Uyttenhove, C. et al. (1988) Proc. Natl. Acad. Sci. USA 85:6934.
    8. Van Snick, J. et al. (1989) J. Exp. Med. 169:363.
    9. Renauld, J-C. et al. (1990) Cytokine 2:9.
    10. Longphre, M. et al. (1999) J. Clin. Invest. 104:1375.
    11. Birner, A. et al. (1992) Exp. Hematol. 20:541.
    12. Holz, L.E. et al. (2005) J. Inflamm. 2:3.
    13. Gounni, A.S. et al. (2000) J. Allergy Clin. Immunol. 106:460.
    14. Olosz, F. and T.R. Malek (2002) J. Biol. Chem. 277:12047.
    15. Bauer, J.H. et al. (1998) J. Biol. Chem. 273:9255.
    16. Jeannin, P. et al. (1998) Blood 91:1355.
    17. Temann, U-A. et al. (2006) Int. Immunol. 19:1.
    18. Poulin, L.F. et al. (2005) Eur. Cytokine Netw. 16:233.
    19. Sitkauskiene, B. et al. (2005) Respir. Res. 6:33.
    20. Gounni, A.S. et al. (2004) J. Immunol. 173:2771.
  • Long Name:
    Interleukin 9
  • Entrez Gene IDs:
    3578 (Human); 16198 (Mouse)
  • Alternate Names:
    Cytokine P40; HP40; IL9; IL-9; IL-9homolog of mouse T cell and mast cell growth factor 40; interleukin 9; interleukin-9; p40 cytokine; p40 T-cell and mast cell growth factor; P40; T-cell growth factor P40
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