Recombinant Human IL-21 GMP Protein, CF

GMP-grade Recombinant Human IL-21 Protein (Catalog # BT-021-GMP) as measured in a cell proliferation assay using B9 mouse hybridoma cells. The ED50 for this effect is 5.00-50.0 ng/mL. Three independent lots were tested foractivity and plotted on the same graph to show lot-to-lot consistency of GMP IL-21.

Equivalent bioactivity of GMP (Catalog # BT-021-GMP) and Animal-Free (BT-021-AFL) grades of Recombinant Human IL-21 as measured in a cell proliferation assay using B9 mouse hybridoma cells (orange and green, respectively).

2 μg/lane of Recombinant Human IL-21 GMP Protein (Catalog # BT-021-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 15-17 kDa, under reducing conditions.

Donor PBMCs were grown using ExCellerate NK Cell Expansion Media (Catalog # CCM037) supplemented with IL-2 (Catalog # BT-002-GMP), IL-12 (Catalog # 10018-IL), IL-15 (Catalog # BT-015-GMP), IL-18 (Catalog # BT-018-GMP), and IL-21 (Catalog # BT-021-GMP) on plates coated with anti-human NKp46 (Catalog # MAB1850). PBMCs were plated to contain 1.8e4 NK cells as determined by flow cytometry on day 0. On day 5, 0.1e6 total cells were taken to seed a 6 well plate. Viability, expansion, and NK purity were analyzed on day 9, with n=5 +/- SEM. A: Total expansion of CD56+ CD3- cells between day 0 and day 9. B: PBMCs grown in ExCellerate NK Cell Expansion Media supplemented with cytokines exhibit viability greater than 80% at day 9.

Donor PBMCs were grown using ExCellerate NK Cell Expansion Media (Catalog # CCM037) supplemented with IL-2 (Catalog # BT-002-GMP), IL-12 (Catalog # 10018-IL), IL-15 (Catalog # BT-015-GMP), IL-18 (Catalog # BT-018-GMP), and IL-21 (Catalog # BT-021-GMP) on plates coated with anti-human NKp46 (Catalog # MAB1850). PBMCs were plated to contain 1.8e4 NK cells as determined by flow cytometry on day 0. On day 5, 0.1e6 total cells were taken to seed a 6 well plate. Viability, expansion, and NK purity were analyzed on day 9, with n=5 +/- SEM for day 9; n=1 for day 0. A: Purity of NK cells at day 9. B: NK ExCellerate media preferentially expands CD56+ NK cells. Percentage of CD56+ cells in culture rose from less than 10% in both donors to more than 60% by day 9.

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

Recombinant Human IL-21 GMP Protein, CF Summary

• Details of Functionality: Measured in a cell proliferation assay using B9 mouse hybridoma cells. The ED₅₀ for this effect is 5.00-50.0 ng/mL.
  The specific activity of recombinant human IL-21 is >1.00 x 10⁶ units/mg, which is calibrated against an internal reference standard for human IL-21.
• Source: E. coli-derived human IL-21 protein
  Gln32-Ser162 with a N-terminal Met
  Produced using non-animal reagents in an animal-free laboratory.
  Manufactured and tested under cGMP guidelines.
• Accession #: Q9HBE4.3
• N-terminal Sequence: Met-Gln32-Asp-Arg-His-Met-Ile-Arg-Met-Arg
• Protein/Peptide Type: GMP Recombinant Proteins
• Purity: >95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 15396 Da ± 50 Da
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 15 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: 15-17 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: >95%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 15396 Da ± 50 Da
• Reconstitution Instructions: Reconstitute the 25 μg size at 250 μg/mL in PBS. Reconstitute all other sizes at 500 μg/mL in PBS.

Notes

Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human IL-21 GMP Protein, CF

• CVID11
• IL21
• IL-21
• IL-21Za11interleukin-21
• interleukin 21
• interleukin-21 isoform
• Za11

Background

Interleukin-21 (IL-21) is an approximately 14 kDa four-helix-bundle member of the family of cytokines that utilize the common gamma chain ( gamma _c) as a receptor subunit. gamma _c is also a subunit of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15 (1). IL-21 is produced by activated T follicular helper cells (Tfh), Th17 cells, and NKT cells (2-6). It exerts its biological effects through a heterodimeric receptor complex of gamma _c and the IL-21-specific IL-21 R (2, 7). Tfh-derived IL-21 plays an important role in the development of humoral immunity through its autocrine effects on the Tfh cell and paracrine effects on immunoglobulin affinity maturation, plasma cell differentiation, and B cell memory responses (4, 8, 9). It is also required for the migration of dendritic cells to draining lymph nodes (10). IL-21 regulates several aspects of T cell function. It co-stimulates the activation, proliferation, and survival of CD8⁺ T cells and NKT cells and promotes Th17 cell polarization (3, 5, 6, 11, 12). It blocks the generation of regulatory T cells and their suppressive effects on CD4⁺ T cells (13, 14). IL-21 R engagement enhances the cytolytic activity and IFN-gamma production of activated NK cells but limits the expansion of resting NK cells (15). In addition, IL-21 suppresses cutaneous hypersensitivity reactions by limiting allergen-specific IgE production and mast cell degranulation (16). Dysregulation of the IL-21/IL-21 R system contributes to the development of multiple immunological disorders (1, 17). The 133 amino acid (aa) mature human IL-21 shares 63% and 61% aa sequence identity with mouse and rat IL-21, respectively. Alternative splicing generates an additional isoform with a substitution of the C-terminal 16 amino acids (18).

1. Tangye, S.G. (2015) Curr. Opin. Immunol. 34:107.
2. Parrish-Novak, et al. (2000) Nature 408:57.
3. Coquet, J.M. et al. (2007) J. Immunol. 178:2827.
4. Vogelzang, A. et al. (2008) Immunity 29:127.
5. Korn, T. et al. (2007) Nature 448:484.
6. Nurieva, R. et al. (2007) Nature 448:480.
7. Asao, H. et al. (2001) J. Immunol. 167:1.
8. Zotos, D. et al. (2010) J. Exp. Med. 207:365.
9. Rankin, A.L. et al. (2011) J. Immunol. 186:667.
10. Jin, H. et al. (2009) J. Clin. Invest. 119:47.
11. Frohlich, A. et al. (2009) Science 324:1576.
12. Yi, J.S. et al. (2009) Science 324:1572.
13. Peluso, I. et al. (2007) J. Immunol. 178:732.
14. Bucher, C. et al. (2009) Blood 114:5375.
15. Kasaian, M.T. et al. (2002) Immunity 16:559.
16. Tamagawa-Mineoka, R. et al. (2011) J. Invest. Dermatol. 131:1513.
17. Ma, J. et al. (2011) Cytokine 56:133.
18. Rahman, M. et al. (2007) FEBS Lett. 581:4001.

Bioinformatics

• Uniprot:
  • Q9HBE4.3()