Recombinant human IL-3 protein (Qk090)

Recombinant Human Interleukin 3 (IL-3) is a pleiotropic cytokine which plays a crucial role in hematopoiesis and acts as a multi-lineage growth factor.  As a hematopoietic growth factor, it promotes the proliferation, differentiation, and survival of various hematopoietic stem cells towards myeloid progenitors. IL-3 is commonly used in cell culture to stimulate the differentiation and maturation of human induced pluripotent stem cells towards mast cells, basophils, neutrophils, eosinophils, monocytes, and megakaryocytes.

Human IL-3 comprises 140 amino acids with a molecular weight of 15.2 kDa. This protein is animal origin-free, carrier protein-free, and tag-free to ensure a homogenous population with exceptional lot-to-lot consistency. IL-3 is suitable for reproducible and high-quality myeloid progenitors and more specific lineages.

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1000µg will be despatched as 2 x 500µg

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Summary

  • Highly pure human protein (UniProt number: P08700)

  • >98%, by SDS-PAGE quantitative densitometry

  • Source: Expressed in E. coli

  • 15.2 kDa monomer

  • Animal origin-free (AOF) and carrier protein-free

  • Manufactured in Cambridge, UK

  • Lyophilized from acetonitrile/TFA

  • Resuspend in 10 mM HCl, prepare single-use aliquots, add carrier protein if desired and store frozen at -20°C or -80°C

Featured applications

  • Generation of iPSC-derived myeloid progenitors

  • Maintenance of peripheral blood cells

  • Proliferation of hematopoietic stem cells

  • Differentiation of peripheral blood cells to myeloid lineages

  • Differentiation of myeloid cells into antigen-presenting cells/monocytes

  • Differentiation of myeloid progenitors into erythrocytes and megakaryocytes

Multilineage colony-stimulating factor; Hematopoietic growth factor; P-cell stimulating factor; Mast-cell growth factor; Colony-stimulating factor

human

species similarity:
mouse – 27%
rat – 26%
bovine – 30%

Bioactivity

Bioactivity graph showing the EC50 of 102 pg/ml (6.7 pM) for Qkine recombinant IL-3

IL-3 activity is determined using proliferation of TF-1 human myeloid leukemia cells. EC50 = 102 pg/ml (6.7 pM). Cells are treated in triplicate with a serial dilution of IL-3 for 72 hours. Cell viability is measured using the CellTiter-Glo (Promega) luminescence assay. Data from Qk090 lot #204548.

Purity

SDS-PAGE gel showing the high purity reduced and non-reduced forms of IL-3

Recombinant IL-3 migrates as a major band at approximately 15.2 kDa in non-reducing (NR) and reduced (R) conditions. No contaminating protein bands are present. The purified recombinant protein (3 µg) was resolved using 15% w/v SDS-PAGE in reduced (+β-mercaptoethanol, R) and non-reduced conditions and stained with Coomassie Brilliant Blue R250. Data from Qk090 batch #204548.

Further quality assays

  • Mass spectrometry, single species with the expected mass

  • Endotoxin: <0.005 EU/μg protein (below the level of detection)

  • Recovery from stock vial: >95%

We are a company founded and run by scientists to provide a service and support innovation in stem cell biology and regenerative medicine.  All our products are exceptionally high purity, with complete characterisation and bioactivity analysis on every lot.

Qkine IL-3 is as biologically active as the comparable alternative supplier protein

Stimulation of proliferation of TF-1 cells with Qkine IL-3 (Qk090, green) and alternative supplier IL-3 (Supplier B, black). Cells were treated in triplicate with a serial dilution of IL-3 for 72 hours and proliferation measured using the CellTiter-Glo (Promega) luminescence assay.

Protein background

Interleukin 3 (IL-3) also known as B-cell stimulatory factor 2/interferon beta 2, is a pleiotropic cytokine with a crucial role in hematopoiesis and multi-lineage growth factor [1–4]. It promotes the proliferation, differentiation, and survival of hematopoietic stem cells and myeloid progenitors [3–6]. IL-3 induces the development of mast cells, basophils, monocytes, erythrocytes, and megakaryocytes and mediates the release of mediators from these cells. It is also involved in the recruitment of myeloid cells to inflammation sites and regulating T-cell function [2].

IL-3 is a monomer glycoprotein composed of 140 amino acids with a molecular weight of approximately 15.7 kDa [7]. The primary structure of IL-3 includes four alpha helices and a bundle of beta sheets stabilized by disulfide bridges [8]. It is produced by various cells including activated T cells, B cells, macrophages, monocytes, mast cells, vascular endothelial cells, and fibroblasts [5]. IL-3 binds with high affinity to the IL-3 receptor (IL-3R) of the hematopoietic receptor superfamily expressed on hemopoietic progenitor cells, monocytes, and B lymphocytes. Ligand stimulation of IL-3R induces tyrosine phosphorylation of effector proteins which subsequently trigger a cascade of intracellular responses such as the PI3k/Akt, Ras/ERK, and MAPK pathways [9,10]. IL-3R is heterodimeric and is composed of two subunits: an alpha subunit (IL-3Rα) and a beta subunit (βc) [11]. IL-3Rα is specific to IL-3, while βc is shared with other cytokine receptors, including those for IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF) [5,10,12].

IL-3 is used in cell culture for the maintenance of peripheral blood cells13 and to induce the expansion and differentiation of hematopoietic stem cells [14]. IL-3 acts synergistically with IL-6 to enhance the proliferation of hemopoietic progenitors1. IL-3 is also used as an early regulator for the differentiation of induced pluripotent stem cells into myeloid and more specific lineages using other lineage instructive cytokines [14,15]. For example, antigen-presenting cells can be derived with the addition of GM-CSF and IL-4 for dendritic cells or M-CSF for macrophages [15]. Finally, IL-3 is also used as a growth factor for the differentiation of erythrocytes with IL-12 and megakaryocytes with IL-6 [16,17].

IL-3 has been studied for conditions which could benefit from the production of early myeloid progenitors and hematopoietic recovery [2]. These include bone marrow reconstitutions, aplastic anemias, and bone marrow disorders such as acute myeloid leukemia and myelodysplastic syndromes [2,18]. Research into the use of IL-3 in immunotherapy and cancer treatment is ongoing, as it may enhance the anti-tumor immune response. For example, the IL-3 and IL-3R axes could be potential therapeutic targets to treat acute myeloid leukemia [10,18,19]. Finally, the potency of IL-3 in amplifying acute inflammation makes it a potential therapeutic target in sepsis [20].

[1] Ikebuchi, K. et al. Interleukin 6 enhancement of interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc. Natl. Acad. Sci. 84, 9035–9039 (1987).

[2] Ihle, J. N. Interleukin-3 and hematopoiesis. Chem. Immunol. 51, 65–106 (1992).

[3] Schrader, J. W. Interleukin-3. in Growth Factors and Cytokines in Health and Disease (eds. Leroith, D. & Bondy, C.) vol. 2 49–84 (JAI, 1997).

[4] Pixley, F. J. & Stanley, E. R. Chapter 319 – Cytokines and Cytokine Receptors Regulating Cell Survival, Proliferation, and Differentiation in Hematopoiesis. in Handbook of Cell Signaling (Second Edition) (eds. Bradshaw, R. A. & Dennis, E. A.) 2733–2742 (Academic Press, 2010). doi:10.1016/B978-0-12-374145-5.00319-3.

[5] Broughton, S. E. et al. The GM–CSF/IL-3/IL-5 cytokine receptor family: from ligand recognition to initiation of signaling. Immunol. Rev. 250, 277–302 (2012).

[6] Kölle, J. et al. Targeted deletion of Interleukin-3 results in asthma exacerbations. iScience 25, 104440 (2022).

[7] Clark-Lewis, I. et al. Automated Chemical Synthesis of a Protein Growth Factor for Hemopoietic Cells, Interleukin-3. Science 231, 134–139 (1986).

[8] Lindemann, A. & Mertelsmann, R. Interleukin-3: Structure and Function. Cancer Invest. 11, 609–623 (1993).

[9] Peso, L. del, González-Garcı́a, M., Page, C., Herrera, R. & Nuñez, G. Interleukin-3-Induced Phosphorylation of BAD Through the Protein Kinase Akt. Science 278, 687–689 (1997).

[10] Testa, U. et al. Interleukin-3 receptor in acute leukemia. Leukemia 18, 219–226 (2004).

[11] Miyajima, A., Mui, A., Ogorochi, T. & Sakamaki, K. Receptors for granulocyte-macrophage colony-stimulating factor, interleukin-3, and interleukin-5. Blood 82, 1960–1974 (1993).

[12] Hodi, F. S. & Soiffer, R. J. Interleukins. in Encyclopedia of Cancer (Second Edition) (ed. Bertino, J. R.) 523–535 (Academic Press, 2002). doi:10.1016/B0-12-227555-1/00110-6.

[13] Staerk, J. et al. Reprogramming of peripheral blood cells to induced pluripotent stem cells. Cell Stem Cell 7, 20–24 (2010).

[14] Ackermann, M. et al. A 3D iPSC-differentiation model identifies interleukin-3 as a regulator of early human hematopoietic specification. Haematologica 106, 1354–1367 (2020).

[15] Ackermann, M., Dragon, A. C. & Lachmann, N. The Immune-Modulatory Properties of iPSC-Derived Antigen-Presenting Cells. Transfus. Med. Hemotherapy 47, 444–453 (2020).

[16] Ploemacher, R. E., van, S. P., Boudewijn, A. & Neben, S. Interleukin-12 enhances interleukin-3 dependent multilineage hematopoietic colony formation stimulated by interleukin-11 or steel factor. Leukemia 7, 1374–1380 (1993).

[17] Majka, M. B.-K., Jacek Kijowski, Ryan Reca, Janina Ratajczak, Mariusz Z. Ratajczak, Marcin. In vitro expansion of human megakaryocytes as a tool for studying megakaryocytic development and function. Platelets 12, 325–332 (2001).

[18] Munoz, L. et al. Interleukin-3 receptor alpha chain (CD123) is widely expressed in hematologic malignancies. Haematologica 86, 1261–1269 (2001).

[19] Jordan, C. T. et al. The interleukin-3 receptor alpha chain is a unique marker for human acute myelogenous leukemia stem cells. Leukemia 14, 1777–1784 (2000).

[20] Weber, G. F. et al. Interleukin-3 amplifies acute inflammation and is a potential therapeutic target in sepsis. Science 347, 1260–1265 (2015).

Our products are for research use only and not for diagnostic or therapeutic use.  Products are not for resale.

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