Recombinant Human IL-4 GMP Protein, CF

Recombinant Human IL-4 GMP Protein (Catalog # BT-004-GMP) as measured in a cell proliferation assay using TF-1 human erythroleukemic cells. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of Recombinant Human IL-4 GMP Protein.

Equivalent bioactivity of GMP (BT-004-GMP) and Animal-Free (BT-004-AFL) grades of Recombinant Human IL-4 as measured in a cell proliferation assay using TF-1 human erythroleukemic cells (Green & Orange, respectively).

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

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human IL-4 GMP Protein, CF Summary

• Details of Functionality: 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 GMP Recombinant Human IL-4 is >1.0x10⁷ IU/mg, which is calibrated against the human IL-4 WHO International Standard (NIBSC code: 88/656).

• 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.
  Manufactured and tested under cGMP guidelines.
• Accession #: P05112.1
• N-terminal Sequence: Met-His25-Lys-(Cys)-Asp-Ile-Thr-Leu-Gln-Glu
• Protein/Peptide Type: GMP Recombinant Proteins
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 15079 Da ± 10 Da
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 15.1 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 13 kDa, under reducing conditions.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • A minimum of 12 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 15079 Da ± 10 Da
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

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Alternate Names for Recombinant Human IL-4 GMP Protein, CF

• B cell growth factor 1
• BCDF
• B-cell stimulatory factor 1
• BCGF1
• BCGF-1
• binetrakin
• BSF1
• BSF-1
• IL4
• IL-4
• IL-4B_cell stimulatory factor 1
• IL4E12
• interleukin 4
• interleukin-4
• Lymphocyte stimulatory factor 1
• MGC79402
• pitrakinra

Background

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 alpha 1. 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).

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.

Bioinformatics

• Uniprot:
  • P05112.1()