YILIMART - Human IL-15Rα sushi, research gr.(25 μg)

IL-15Rα sushi stands for recombinant human interleukin 15 receptor alpha, soluble sushi domain. It is composed of the extracellular sushi domain of IL-15Rα plus a natural hinge domain, and it is a high affinity agonist of IL-15 in culture, maintaining the natural biological effect of soluble IL-15Rα. Human IL-15Rα sushi is a recombinant protein optimized for use in cell culture, differentiation studies, and functional assays.

Applications

Human IL-15Rα sushi can be used for a variety of applications, including:

In vitro activation and expansion of memory and naive CD8⁺ T cells, NK cells, and NKT cells in combination with IL-15
Study of IL-15/IL-2 receptor signaling
Investigation of new potential adjuvants for vaccine studies

Background information

IL-15Rα is a subunit of the IL-15 receptor together with IL-2Rβ (CD122) and IL-2Rγ (CD132), two subunits shared with IL-2 receptor. A soluble form of IL-15Rα, including the extracellular sushi domain, is naturally released by antigen-presenting cells

in vivo

. Soluble IL-15Rα binds with high affinity IL-15 and acts as IL-15 antagonist through the prevention of IL-15 binding on its α/β/γ receptor. However, on lymphocytes that express only the heterodimeric IL-2Rβ/γ receptor, IL-15Rα presents IL-15

in trans

to the IL-2Rβ/γ receptor, and enhances the effects of IL-15, increasing lymphocyte proliferation and survival. Recombinant Human IL-15Rα sushi is composed of the extracellular sushi domain of IL-15Rα plus a natural hinge domain, and it is a high affinity agonist of IL-15 in culture, maintaining the natural biological effect of soluble IL-15Rα.

Quality description

Research-grade

cytokines are suitable for a wide variety of cell culture applications. They are sterile-filtered prior to lyophilization. Generally, endotoxin levels are <0.1 ng/μg (<1 EU/μg), and purities are >95%. The biological activity is tested in appropriate bioassays.

Premium-grade

cytokines offer the convenience of high and well-defined biological activities and allow exact unit dosing for demanding applications. The biological activity is determined after lyophilization and reconstitution, and normalized to WHO/NIBSC standards whenever available. In general, endotoxin levels are <0.01 ng/μg (<0.1 EU/μg), and purities are >97%. Lot-specific certificates of analysis are available on request (macstec@miltenyibiotec.de).

Biological activity

Proliferation of CTLL-2 cells
premium grade: ≥ 5×

10
⁵
U/mg
research grade: ≥ 3×

10
⁵
U/mg

Figure 1
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Figure 1

Mass spectrometry analysis (ESI-MS) of Human IL-15Rα sushi, premium grade. The peak corresponds to the calculated molecular mass of 9340 Da.

Mass spectrometry analysis (ESI-MS) of Human IL-15Rα sushi, premium grade. The peak corresponds to the calculated molecular mass of 9340 Da.
Figure 2
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Figure 2

IL-15Ra sushi was immobilized to a CM5 chip, and binding kinetics were evaluated with IL-15 concentrations of 500–0.5 nM. Data were analyzed with BIAevaluation software 4.1.1 using an 1:1 model.

Result:
k
_on
(association rate) = 3.7×10
⁵
M
^-1
s
^-1
; k
_off
(dissociation rate) = 4.4×10
^-5
s
^-1
; k
_D
(dissociation constant) = k
_off
/k
_on
= 1.2×10
^-10
M = 120 pM

IL-15Ra sushi was immobilized to a CM5 chip, and binding kinetics were evaluated with IL-15 concentrations of 500–0.5 nM. Data were analyzed with BIAevaluation software 4.1.1 using an 1:1 model.

Result:
k
_on
(association rate) = 3.7×10
⁵
M
^-1
s
^-1
; k
_off
(dissociation rate) = 4.4×10
^-5
s
^-1
; k
_D
(dissociation constant) = k
_off
/k
_on
= 1.2×10
^-10
M = 120 pM
Figure 3
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Figure 3

Human IL-15Rα sushi activity assay.
The biological activity of Human IL-15Rα sushi, premium grade, was determined by inhibition assay using human IL-15-induced proliferation of CTLL-2 cells. Activity of Human IL-15Rα sushi, premium grade, (red line) was compared to another commercially available product (black line).

Human IL-15Rα sushi activity assay.
The biological activity of Human IL-15Rα sushi, premium grade, was determined by inhibition assay using human IL-15-induced proliferation of CTLL-2 cells. Activity of Human IL-15Rα sushi, premium grade, (red line) was compared to another commercially available product (black line).
Figure 4
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Figure 4

SDS-PAGE of Human IL-15Rα sushi, premium grade
under reduced (R) and non-reduced (NR) conditions.

SDS-PAGE of Human IL-15Rα sushi, premium grade
under reduced (R) and non-reduced (NR) conditions.

Selected references
1. Soman, G. et al. (2009) MTS dye based colorimetric CTLL-2 cell proliferation assay for product release and stability monitoring of interleukin-15: assay qualification, standardization and statistical analysis. J. Immunol. Methods 348: 83-94
2. Mortier, E. et al. (2006) Soluble interleukin-15 receptor α (IL-15Rα)-sushi as a selective and potent agonist of IL-15 action through IL-15Rβ/γ: hyperagonist IL-15×IL-15Rα fusion proteins. J. Biol. Chem. 281: 1612-1619
3. Bouchaud, G. et al. (2008) The exon-3-encoded domain of IL-15Rα contributes to IL-15 high-affinity binding and is crucial for the IL-15 antagonist effect of soluble IL-15Rα. J. Mol. Biol. 382(1): 1-12
4. Wu, J. (2013) IL-15 agonists: The cancer cure cytokine. J. Mol. Genet. Med. 7(4): 85
Certificates

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Human IL-15Rα sushi, research gr.(25 μg)

For research use only

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Applications

Background information

Quality description

Biological activity

Figure 1

Figure 1

Figure 2

Figure 2

Figure 3

Figure 3

Figure 4

Figure 4

Selected references

Certificates

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