/, IL-2 family, Recombinant Human Cytokines/Human Interleukin-21 Recombinant

Human Interleukin-21 Recombinant



SKU: RKQ9HBE4 Tags: , , , ,


Optimized DNA sequence encoding Human Interleukin-21 mature chain was expressed in Escherichia Coli.
Molecular weight
Human Interleukin-21 is generated by the proteolytic removal of the signal peptide and propeptide, the molecule has a calculated molecular mass of approximately 15kDa. Recombinant IL-21 is a monomer protein consisting of 132 amino acid residue subunits, and migrates as an approximately 15kDa protein under non-reducing and reducing conditions in SDS-PAGE.
>97%, as determined by SDS-PAGE and HPLC
Biological Activity
The ED(50) was determined by the dose-dependent proliferation of activated B Cells.
Protein Sequence
Endotoxin content was assayed using a LAL gel clot method. Endotoxin level was found to be less than 0.1 ng/µg(1EU/µg).
Recombinant Interleukin-21 was lyophilized from a 0.2 μm filtered PBS solution pH7.5.
A quick spin of the vial followed by reconstitution in distilled water to a concentration not less than 0.1 mg/mL. This solution can then be diluted into other buffers.
The lyophilized protein is stable for at least years from date of receipt at -20° C. Upon reconstitution, this cytokine can be stored in working aliquots at2° -8° C for one month, or at -20° C for six months, with a carrier protein without detectable loss of activity. Avoid repeated freeze/thaw cycles.
This cytokine product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.
Molecular function


Autologous CD4+ T cell secretion of IL-2/IL-21 is necessary but not sufficient to help CD8+ T cells proliferate.

  • IL-2, IL-21, or both were added in each condition.

Mixed lymphocytes tumor cell culture (MLTC)

  • PBMCs isolated from melanoma patients were thawed and incubated overnight at 37°C.
  • PBMCs (106 cells/well) were cultured in the presence of autologous irradiated (150 Gy) melanoma cells at a lymphocyte to tumor ratio of 5:1 in 24-well plates with X-VIVO15 and 5% HS.
  • Different cytokine combinations were added to these MLTCs in order to select the most suitable in vitro culture conditions in terms of T lymphocyte expansion and efficiency in tumor cell recognition: (1) 120 IU/ml rhIL-2 ; (2) 120 IU/ml rhIL-2 and 10 ng/ml rhIL-15 ; (3) 10 ng/ml rhIL-15; (4) 5 ng/ml rh-IL-7 ; (5) 120 IU/ml rhIL-2 and 5 ng/ml rhIL-7; (6) 120 IU/ml rhIL-2 and 10 ng/ml rhIL-21 ; (7) 10 ng/ml rhIL-21 alone.
  • Fresh medium containing the above indicated cytokines was replaced every 3 days.
  • These MLTCs were stimulated weekly with irradiated autologous melanoma cells, and their reactivity was tested starting from the 3rd week of culture.

DC Co-Cultures

  • Sorted naïve (live, singlet, CD3+CD28+CD95-CCR7+CD27+) CD4+ T cells were co-cultured with sorted CD103+ DCs (live, lineage-CD14-HLA-DR+CD11c+) (100:1), sorted CD103− DCs (lineage-CD14-HLA-DR+CD11c+CD103-) (100:1), or anti-CD3/anti-CD28 beads (control, 4:1) in X-Vivo15 media under Th17 conditions (10 U/mL recombinant human IL-2, 12.5 ng/mL rhIL-1β, 25 ng/mL rhIL-21, 25 ng/ml rhIL-23, 10 μg/mL anti-IFNγ, 10 μg/mL anti-IL-12 and 2 ng/mL TGF-β, with or without 25 ng/mL rhIL-6).
  • All cytokines were from , except IL-2, IL-6 and IL-21 .
  • DC co-cultures were stimulated with SEB (1 μg/mL ).
  • All cultures were fed on day 3 with rhIL-2 (2 U/mL), anti-IFNγ (10 μg/mL) and anti-IL-12 (10 μg/mL).
  • After 7 days of co-culture, qRT-PCR was performed for IL17A and RORc gene expression .
  • Gene expression was quantified using ΔΔCT analysis in excel (version 12.3.0).

Decreased CXCR5+ PD1+/CD4+ cell production by dexamethasone.

  • A. Dexamethasone (Dex) reduced the proportion of IL-21 stimulated CXCR5+ PD1+/CD4+ cell.

Identification of IL21, CD40L, αIgM, BAFF and LPS regulated genes in transformed human germinal centre B cells using microarrays.

  • BL2 cell were stimulated with αIgM F(ab)2 fragments (3 hrs) , IL21 (2 hrs) , CD40L (6 hrs) , LPS (6 hrs) and BAFF (9 hrs) .

In vitro Sensitization (IVS) of CD8+ T Cells

  • CTLs were induced as previously described + donors were pulsed with tumor cell lysates for the last 24 h of maturation.
  • Based on the number of tumor cells before lysis, tumor cells were added to DC at a 3∶1 ratio.
  • TA-pulsed mDC were then irradiated (3000 rad) and washed with PBS.
  • Autologous CD8+ T cells were isolated from cryopreserved PBMC by negative selection using magnetic bead separations and added to the mDC at the 10∶1 ratio.
  • Cells were cultured in an atmosphere of 5% CO2 in air at 37°C for 7 days in AIM-V media containing 10 ng/ml IL-7 and 10 ng/ml IL-21 and 5% (v/v) FBS.
  • On day 7, fresh, tumor cell lysate-pulsed and irradiated mDC were added and T cells were cultured for additional 7 days in AIM V containing 20 IU/ml IL-2 , 5 ng/ml IL-7, 10 ng/ml IL-21 and 5% (v/v) FBS.
  • On day 14, cells were…
  • B cells were obtained from peripheral blood of healthy volunteers by CD22 MACS microbead magnetic column selection .
  • Purity was > 97% and was determined by flow cytometry by staining for CD19 and CD20.
  • We co-cultured B cells (2 - 2.5 × 105 cells/ml) on irradiated (50 Gray) CD40L-expressing L cell fibroblasts (5 × 104 cells/ml, > 96% surface CD154+) in 1 ml Iscove's Modified Eagle's Medium containing 8% fetal bovine serum and penicilin/streptomycin and with or without rhIL-4 (25 ng/ml) or rhIL-21 (10 ng/ml) in 24-well plates.
  • After 6 days, we harvested supernatants and the levels of IgG4 in the supernatant were determined as described below.
Stay in the loop
For new product information and regular updates on our product range please fill your details