| Product Description |
Recombinant HEK293 cells
stably express STAT6-RE-Luc and human interleukin 4
Receptor (IL-4R)
on the surface and activate the receptor to the downstream signaling pathways
upon the binding of IL-4 and IL-13. The surface expression of IL-4R
is validated by luminescence analysis. This stable cell line product is
designed for cell-based functional assays that measure binding affinity and
stability of antibody based biologics binding with IL-4 and IL-13.
|
| Applications |
Assessment of
bioactivity of IL-4 and IL-13.
|
| Host Cell |
HEK293
|
| Size |
2 vials of frozen
cells (>1X106 per
vial in 1 ml)
|
| Storage |
Store
cells in liquid nitrogen immediately upon receipt. Thaw and recover cells
within one year from the date received.
|
| Mycoplasma Status |
Negative. The mycoplasma
test was performed with MycoAlert™ PLUS Mycoplasma Detection Kit (Cat. No.
LT07-318, Lonza).
|
| Stability |
Stable through more
than 15 passages with no significant changes in assay performance or expression
profile.
|
| Culture Properties |
Adherent
|
| Freeze Medium |
42%
DMEM (Cat. No.10569-010, Gibco), 50% FBS (Cat. No.10099-141, Gibco), 8% DMSO
(Cat. No. D2650, Sigma)
|
| Complete Growth Medium |
DMEM,
10% FBS
|
| Culture Medium |
DMEM,
10% FBS, 8 μg/ml Puromycin (Cat. No.A11138-02, Gibco), 300 μg/ml Geneticin
(Cat. No.10131-027, Gibco), 200 μg/ml Hygromycin B (Cat. No. 10687010,
Invitrogen), 100 μg/ml Zeocin (Cat. No. R250-01, Life Technologies)
|
| Background |
IL-4Ralpha together
with IL-13R alpha, can form a functional receptor for IL-4; can be also one of
the functional forms of the IL-13 receptor. IL-4R/IL-13R can bind interleukin 4
and interleukin 13 to regulate IgE production. IL-4R alpha also can bind
interleukin 4 to promote differentiation of Th2 cells. A soluble form of the
encoded protein can be produced by proteolysis of the membrane-bound protein,
and this soluble form can inhibit IL4-mediated cell proliferation and IL-5
upregulation by T-cells. Allelic variations in IL-4R alpha have been associated
with atopy, a condition that can manifest itself as allergic rhinitis, sinusitus,
asthma, or eczema. Polymorphisms in this gene are also associated with
resistance to human immunodeficiency virus type-1 infection.
|
Figure 1. Luminescence analysis of activation of STAT6-RE-Luc and IL-4R on HEK293 cells. The HEK293/STAT6-RE-Luc cells were stimulated by IL-4 (Cat. No. Z02925-50, Genscript). Fire-Lumi™ Reagent (Cat. No. L00877C-1000, Genscript) was added and luminescence was measured by PHERAstar FSX.
Figure 2. Luminescence analysis of activation of STAT6-RE-Luc and IL-13R on HEK293 cells. The HEK293/STAT6-RE-Luc cells were stimulated by IL-13 (Cat. No. Z03020-50, Genscript). Fire-Lumi™ Reagent (Cat. No. L00877C-1000, Genscript) was used and luminescence measured by PHERAstar FSX.
Notes:
EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom) / (1+10^ ((LogEC50-X)*Hill Slope))
X is the logarithm of concentration. Y is the response.
Y is RLU and starts at Bottom and goes to Top along a sigmoid curve.
For
research use only. Not intended for human and animal therapeutic or diagnostic
use.
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