Rat IL-3 Antibody

Catalog #: AF2524 Datasheet

Catalog #	Availability	Size / Price	Qty
AF2524
AF2524-SP

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Rat IL-3 Antibody Summary

Species Reactivity

Rat

Specificity

Detects rat IL-3 in direct ELISAs and Western blots. In direct ELISAs and Western blots, approximately 5% cross-reactivity with recombinant mouse IL-3 and less than 1% cross-reactivity with recombinant human IL-3 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant rat IL‑3
Ile27-Cys169
Accession # P97688

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‑3 (Catalog # 2524-RL)

Immunocytochemistry

5-15 µg/mL

See below

Neutralization

Measured by its ability to neutralize IL‑3-induced proliferation in the M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line. Holmes, K.L. et al. (1985) Proc. Natl. Acad. Sci. USA 82:6687. The Neutralization Dose (ND₅₀) is typically 0.75-3 µg/mL in the presence of 0.5 ng/mL Recombinant Rat IL‑3.

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

Immunocytochemistry

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IL‑3 in Rat Splenocytes. IL-3 was detected in immersion fixed rat splenocytes using Goat Anti-Rat IL-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2524) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (yellow; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Neutralization

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Cell Proliferation Induced by IL‑3 and Neutralization by Rat IL‑3 Antibody. Recombinant Rat IL‑3 (Catalog # 2524-RL) stimulates proliferation in the M‑NFS‑60 mouse myelogenous leukemia lymphoblast cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Rat IL‑3 (0.5 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Rat IL‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2524). The ND₅₀ is typically 0.75-3 µg/mL.

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-3

Rat interleukin-3 (IL-3; also multi-CSF) is a 26 kDa, variably glycosylated monomeric polypeptide that belongs to the alpha -helix family of hematopoietic cytokines (1, 2). IL-3 has pleiotrophic activies on a number of hematopoietic-related cells (1, 3). The rat molecule has two alternate splice forms. The first is termed IL-3 beta and is synthesized as a 169 amino acid (aa) precursor that contains a 27 aa signal sequence and a 142 aa mature segment (1, 2). The second is called IL-3 alpha, and is identical to IL-3 beta, save for a three amino acid (Tyr-Pro-Gln) deletion at positions 56-58 (1). The beta form is considered the most common form. Each form has an alpha -helical structure with two intrachain disulfide bonds and two potential N-linked glycosylation sites. Rat IL-3 is generally considered to be species-specific in its activity. In the mature region, rat IL-3 shares 55%, 30%, and 24% aa sequence identity with mouse, human, and bovine IL-3, respectively. Cells known to express IL-3 include connective tissue mast cells, astrocytes, microglia, megakaryocytes, eosinophils, T cells, keratinocytes and thymic epithelium.

IL-3 exerts its biological activities by binding to a 70 kDa, low affinity, ligand-binding IL-3 R alpha subunit, (6) which then recruits a 120 kDa, common beta -chain, signal transducing subunit (7) to form a functional IL-3 receptor (1, 6, 7). Receptors for IL-3 are present on bone marrow progenitors, macrophages, mast cells, eosinophils, megakaryocytes, basophils, and various myeloid leukemic cells. IL-3 can stimulate the proliferation and differentiation of pluripotent hematopoietic stem cells as well as various lineage committed progenitors including those for neutrophils, macrophages, magakaryocytes, and erythroid cells. IL-3 can stimulate the growth of early B cells and mature macrophages, mast cells, eosinophils, basophils, and megakaryocytes. IL-3 augments the function activity of basophils, mast cells, eosinophils, and macrophages (1, 8). In combination with other molecules such as CD40L and or IL-4, IL-3 can stimulate production of dendritic cells (1, 2, 9, 10).

References

Martinez-Moczygemba, M. and D.P. Huston (2003) J. Allergy Clin. Immunol. 112:653.
Mangi, M.H. and A.C. Newland (1999) Cytokines Cell. Mol. Ther. 5:87.
Esandi, M. del C. et al. (1998) Gene 211:151.
Cohen, D.R. et al. (1986) Nucleic Acids Res. 14:3641.
Gebicke-Haerter, P.J. et al. (1994) J. Neuroimmunol. 50:203.
Chritton, S.C. and M.H. Sheng (1997) GenBank Accession # NP_640353.
Appel, K. et al. (1995) J. Neurosci. 15:5800.
Schrader, J.W. (2001) in Cytokine Reference, Oppenhiem, J.J. and M. Feldmann (eds): Academic Press, p. 855.
Ebner, S. et al. (2002) J. Immunol. 168:6199.
Buelens C. et. al. (2002) Blood 99:993.

Long Name

Interleukin 3

Entrez Gene IDs

3562 (Human); 16187 (Mouse); 24495 (Rat)

Alternate Names

Hematopoietic growth factor; IL3; IL-3; IL-3MGC79398; interleukin 3 (colony-stimulating factor, multiple); interleukin-3; Mast cell growth factor; mast-cell growth factor; MCGF; MCGFMGC79399; MULTI-CSF; multilineage-colony-stimulating factor; Multipotential colony-stimulating factor; P-cell stimulating factor; P-cell-stimulating factor