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Recombinant Human IL-4 Protein, Animal-Free

Bio-Techne includes R&D Systems | Catalog # BT-004-AFL

Intended for preclinical researchers who may transition to GMP IL-4 for their clinical work
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BT-004-AFL-01M
BT-004-AFL-050

Key Product Details

Source

E. coli

Accession #

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

E. coli-derived human IL-4 protein
His25-Ser153, with an N-terminal Met
Produced using non-animal reagents in an animal-free laboratory.

Purity

>97%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

15.1 kDa

SDS-PAGE

13 kDa, under reducing conditions.

Activity

Measured in a cell proliferation assay using TF‑1 human erythroleukemic cells. Kitamura, T. et al. (1989) J. Cell Physiol. 140:323.
The ED50 for this effect is 0.0400-0.320 ng/mL.
The specific activity of Animal-Free™ Recombinant Human IL-4 is >1.0x10IU/mg, which is calibrated against the human IL-4 WHO International Standard (NIBSC code: 88/656).

Scientific Data Images for Recombinant Human IL-4 Protein, Animal-Free

Recombinant Human IL-4 Protein Bioactivity Comparison.

As an alternative, please consider our next generation Recombinant Human IL-4 Protein, (BT-004-GMP). This has equivalent bioactivity to Recombinant Human IL-4 Proteins (BT-004) and Catalog # BT-004-AFL.These combine R&D Systems quality with scalability that allows for a solid supply chain. All Recombinant Human IL-4 proteins are measured in a cell proliferation assay using TF-1 human erythroleukemic cell line.

Animal-Free™ Recombinant Human IL‑4 Protein Bioactivity.

Recombinant Human IL‑4 stimulates cell proliferation of TF‑1 human erythroleukemic cells. The ED50 for this effect is 0.0400‑0.320 ng/mL.

Animal-Free™ Recombinant Human IL‑4 Protein SDS-PAGE.

2 μg/lane of Animal-Free™ Recombinant Human IL‑4 Protein (Catalog # BT-004-AFL) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 13 kDa underreducing conditions.

Formulation, Preparation and Storage

BT-004-AFL
Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
Reconstitution Reconstitute at 500 μg/mL in PBS. 
Shipping The product is shipped at ambient temperature. Upon receipt, store it 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.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: IL-4

Interleukin-4 (IL-4), also known as B cell-stimulatory factor-1, is a monomeric, approximately 13 kDa‑18 kDa Th2 cytokine that shows pleiotropic effects during immune responses (1‑3). It is a glycosylated polypeptide that contains three intrachain disulfide bridges and adopts a bundled four alpha-helix structure (4). Human IL-4 is synthesized with a 24 aa signal sequence. Alternate splicing generates an isoform with a 16 aa internal deletion. Mature human IL-4 shares 55%, 39% and 43% aa sequence identity with bovine, mouse, and rat IL-4, respectively. Human, mouse, and rat IL-4 are species-specific in their activities (5‑7). IL-4 exerts its effects through two receptor complexes (8, 9). The type I receptor, which is expressed on hematopoietic cells, is a heterodimer of the ligand binding IL-4 R alpha and the common gamma chain (a shared subunit of the receptors for IL-2, -7, -9, -15, and ‑21). The type II receptor on nonhematopoietic cells consists of IL-4 R alpha and IL‑13 R alpha1. The type II receptor also transduces IL-13 mediated signals. IL-4 is primarily expressed by Th2-biased CD4+ T cells, mast cells, basophils, and eosinophils (1, 2). It promotes cell proliferation, survival, and immunoglobulin class switch to IgG4 and IgE in human B cells, acquisition of the Th2 phenotype by naïve CD4+ T cells, priming and chemotaxis of mast cells, eosinophils, and basophils, and the proliferation and activation of epithelial cells (10‑13). IL-4 plays a dominant role in the development of allergic inflammation and asthma (12, 14).

Due to its role in the differentiation of certain immune cell types, IL-4 is commonly used in combination with other growth factors to transform induced pluripotent stem cells into dendritic cells in high numbers. These dendritic cells can then be used for research or clinical applications to improve disease modeling, for screening and cell therapies (15). Study of IL-4 signaling has led to the development of monoclonal antibodies that can block the signaling pathway at various steps to mitigate the inflammatory response in certain autoimmune diseases (16, 17). While IL-4 has the capacity to improve immune functions, treatments involving IL-4 have not been utilized due to the dangerous side effects that may result from IL-4 signaling in non-immune cells (16). Blockade of IL-4 signaling also has been studied as a therapeutic target to suppress inflammation in the tumor microenvironment (18). Use of IL-4 suppressors can also improve the efficacy of anti-tumor immunotherapies, as blocking IL-4 enhances activity of tumor-specific T lymphocytes (19, 20).

References

  1. Benczik, M. and S.L. Gaffen (2004) Immunol. Invest. 33:109.
  2. Chomarat, P. and J. Banchereau (1998) Int. Rev. Immunol. 17:1.
  3. Yokota, T. et al. (1986) Proc. Natl. Acad. Sci. 83:5894.
  4. Redfield, C. et al. (1991) Biochemistry 30:11029.
  5. Ramirez, F. et al. (1988) J. Immunol. Meth. 221:141.
  6. Leitenberg, D. and T.L. Feldbush (1988) Cell. Immunol. 111:451.
  7. Mosman, T.R. et al. (1987) J. Immunol. 138:1813.
  8. Mueller, T.D. et al. (2002) Biochim. Biophys. Acta 1592:237.
  9. Nelms, K. et al. (1999) Annu. Rev. Immunol. 17:701.
  10. Paludan, S.R. (1998) Scand. J. Immunol. 48:459.
  11. Corthay, A. (2006) Scand. J. Immunol. 64:93.
  12. Ryan, J.J. et al. (2007) Crit. Rev. Immunol. 27:15.
  13. Grone, A. (2002) Vet. Immunol. Immunopathol. 88:1.
  14. Rosenberg, H.F. et al. (2007) J. Allergy Clin. Immunol. 119:1303.
  15. Flosdorf, N. & Zenke, M. (2022) Eur. J. Immunol. 52:1880.
  16. Junttila, I.S. (2018) Front Immunol. 9:888.
  17. Keegan, A.D. et al. (2021) Fac Rev. 10:71.
  18. Bankaitis, K.V. & Fingleton, B. (2016) Clin. Exp. Metastasis. 32:847.
  19. Mirlekar, B. (2022) SAGE Open Med. 10:1.
  20. Ilto, S. E. et al. (2017) Cancer Immunol Immunother. 66:1485.

Long Name

Interleukin 4

Alternate Names

BCDF, BCGF-1, BCGF1, BSF-1, BSF1, IL4, IL4E12

Entrez Gene IDs

3565 (Human); 16189 (Mouse); 287287 (Rat); 397225 (Porcine); 280824 (Bovine); 403785 (Canine); 574281 (Primate); 100302454 (Rabbit)

Gene Symbol

IL4

UniProt

Additional IL-4 Products

Product Documents for Recombinant Human IL-4 Protein, Animal-Free

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.

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.  

 Production

  • All molecular biology procedures use animal-free media and dedicated labware.
  • Dedicated fermentors are utilized in committed animal-free areas.

Purification

  • Protein purification columns are animal-free.
  • Bulk proteins are filtered using animal-free filters.
  • Purified proteins are stored in animal-free containers in a dedicated cold storage room.

    Quality Assurance

    • Low Endotoxin Level.
    • No impairment of biological activity.
    • High quality product obtained under stringent conditions.
    • For ex vivo research or bioproduction, additional documentation can be provided.

    Please read our complete Animal-Free Statement

    Product Specific Notices for Recombinant Human IL-4 Protein, Animal-Free

    For research use or further manufacturing only

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