Key Product Details

Validated by

Biological Validation

Species Reactivity

Rat

Applications

Western Blot, Neutralization, Intracellular Staining by Flow Cytometry, CyTOF-ready

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all IL-9 Primary Antibodies »

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant rat IL‑9
Gln19-Ala144
Accession # NP_001099217

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Scientific Data Images for Rat IL‑9 Antibody

Cell Proliferation Induced by IL‑9 and Neutralization by Rat IL‑9 Antibody.

Cell Proliferation Induced by IL‑9 and Neutralization by Rat IL‑9 Antibody.

Recombinant Rat IL-9 (Catalog # 4134-RL) stim-ulates 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.

Detection of IL-9 antibody in Rat Spleno-cytes antibody by Flow Cytometry.

Detection of IL‑9 in Rat Spleno-cytes 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 permea-bilized with saponin.

Applications for Rat IL‑9 Antibody

Application
Recommended Usage

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
Sample: Mouse splenocytes treated with PMA and Ca2+ ionomycin, fixed with paraformaldehyde, and permeabilized with saponin

Western Blot

0.1 µg/mL
Sample: Recombinant Rat IL‑9 (Catalog # 4134-RL)

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.

Flow Cytometry Panel Builder

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Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Calculators

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.

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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

Alternate Names

IL9

Entrez Gene IDs

3578 (Human); 16198 (Mouse)

Gene Symbol

IL9

UniProt

NP_001099217

Additional IL-9 Products

Product Documents for Rat IL‑9 Antibody

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Product Specific Notices for Rat IL‑9 Antibody

For research use only

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