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Rat IL-9 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF4134

R&D Systems, part of Bio-Techne
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AF4134
AF4134-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Rat

Applications

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

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG

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.

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.

Reconstitution Buffer Available:
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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.

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

Additional IL-9 Products

Product Documents for Rat IL-9 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Rat IL-9 Antibody

For research use only

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