Rat IL-9 Antibody

Catalog #: AF4134 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF4134
AF4134-SP

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

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Rat IL-9 Antibody Summary

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 either lyophilized or as a 0.2 µm filtered solution in PBS.

Endotoxin Level

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

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.

Scientific Data

Neutralization

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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.

Intracellular Staining by Flow Cytometry

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

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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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile 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

Renauld, J-C. et al. (1993) Adv. Immunol. 54:79.
Demoulin J-B. and J-C. Renauld (1998) Intern. Rev. Immunol. 16:345.
Knoops, L. and J-C. Renauld (2004) Growth Factors 22:207.
Finiasz, M.R. et al. (2006) Clin. Exp. Immunol. 147:139.
Flubacher, M.M. et al. (1994) J. Virol. 68:7709.
Blanco, J.C. et al. (2004) J. Interferon Cytokine Res. 24:21.
Uyttenhove, C. et al. (1988) Proc. Natl. Acad. Sci. USA 85:6934.
Van Snick, J. et al. (1989) J. Exp. Med. 169:363.
Renauld, J-C. et al. (1990) Cytokine 2:9.
Longphre, M. et al. (1999) J. Clin. Invest. 104:1375.
Birner, A. et al. (1992) Exp. Hematol. 20:541.
Holz, L.E. et al. (2005) J. Inflamm. 2:3.
Gounni, A.S. et al. (2000) J. Allergy Clin. Immunol. 106:460.
Olosz, F. and T.R. Malek (2002) J. Biol. Chem. 277:12047.
Bauer, J.H. et al. (1998) J. Biol. Chem. 273:9255.
Jeannin, P. et al. (1998) Blood 91:1355.
Temann, U-A. et al. (2006) Int. Immunol. 19:1.
Poulin, L.F. et al. (2005) Eur. Cytokine Netw. 16:233.
Sitkauskiene, B. et al. (2005) Respir. Res. 6:33.
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