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Rat IL‑3 Antibody

Bio-Techne includes R&D Systems | Catalog # AF2524

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AF2524
AF2524-SP

Key Product Details

Species Reactivity

Rat

Applications

Immunocytochemistry, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Summary for Rat IL‑3 Antibody

Immunogen

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

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Rat IL‑3 Antibody

IL-3 antibody in Rat Splenocytes by Immunocytochemistry (ICC).

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.
Cell Proliferation Induced by IL‑3 and Neutralization by Rat IL‑3 Antibody.

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.

Applications for Rat IL‑3 Antibody

Application
Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed rat splenocytes

Western Blot

0.1 µg/mL
Sample: Recombinant Rat IL‑3 (Catalog # 2524-RL)

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 of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Endotoxin Level

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

Reconstitution

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

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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

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-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

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

Long Name

Interleukin 3

Alternate Names

IL3, MCGF, MULTI-CSF

Entrez Gene IDs

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

Gene Symbol

IL3

UniProt

Additional IL-3 Products

Product Documents for Rat IL‑3 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‑3 Antibody

For research use only

Reconstitution Calculator

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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