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CYTOKINE SIGNALING 4
108
HEK-Blue IL-4/IL-13 Cells - IL-4 and IL-13 Reporter Cells
➥ Monitor the JAK/STAT-6 pathway
➥ Detection of human IL-4 and human/murine IL-13
Background
The transcription factor STAT6 is activated primarily by two cytokines
with overlapping biologic functions, IL-4 and IL-13. It can also be activated
by IFN-a in a cell-specific manner. In non-hematopoietic cells, IL-4 and
IL-13 bind a receptor complex composed of the IL-4Ralpha and
IL-13Ralpha1. Upon ligand binding, the receptor complex activates the
receptor-associated Janus kinases (JAK1 and Tyk2) leading to the
recruitment of STAT6 and its phosphorylation. Activated STAT6 forms
homodimers that translocate to the nucleus where they bind the
promoter of responsive genes inducing gene transcription.
Description
HEK-Blue IL-4/IL-13 cells allow the detection of bioactive IL-4 and
IL-13 by monitoring the activation of the STAT-6 pathway. These cells
were generated by stable transfection of HEK293 cells with the human
STAT6 gene to obtain a fully active STAT6 pathway. The other genes of
the pathway are naturally expressed in sufficient amounts. The cells were
further transfected with a SEAP reporter gene under the control of the
IFNb minimal promoter fused to four STAT6 binding sites. HEK-Blue
IL-4/IL-13 cells produce SEAP in response to IL-4 or IL-13 stimulation
and to a lower extent IFN-a. The levels of SEAP secreted in the
supernatant can be easily determined with QUANTI-Blue , a SEAP
detection medium (Figure 2).
Stimulation of HEK-Blue IL-4/IL-13 cells with IL-4 or IL-13 can be blocked
by the neutralizing monoclonal anti-hIL-4-IgA and anti-hIL-13-IgA
antibodies, respectively (see page 104).
• Detection range for IL-4: 0.5 - 100 ng/ml
• Detection range for IL-13: 5 - 1000 ng/ml
HEK-Blue IL-4/IL-13 cells are resistant to blasticidin and Zeocin .
Contents and Storage
HEK-Blue IL-4/IL-13 cells are grown in standard DMEM medium, 2 mM
L-glutamine with 10% FBS supplemented with blasticidin (10 μg/ml) and
Zeocin (100 μg/ml). Each vial contains 5-7 x 10 6 cells and is supplied
with 100 μl of blasticidin at 10 mg/ml, 100 μl of Zeocin at 100 mg/ml,
1 ml Normocin at 50 mg/ml and 1 pouch of QUANTI-Blue . Cells are
shipped on dry ice.
HEK-Blue IL-4/IL-13 cells are also available in a kit. See below:
HEK-Blue IL-4/IL-13 Kit
• HEK-Blue IL-4/IL-13 Cells 5-7 x 10 6 cells
• Anti-hIL-4-IgA antibody 100 μg
• Anti-hIL-13-IgA antibody 100 μg
• QUANTI-Blue 1 pouch (100 ml)
• Normocin 1 ml (50 mg/ml)
• Blasticidin 100 μl (10 mg/ml)
• Zeocin 100 μl (100 mg/ml)
www.invivogen.com/cytokine-sensor-cells
Figure 1: JAK/STAT6 signaling pathway induced by IL-4 and
IL-13.
Figure 2: Stimulation of HEK-Blue IL-4/IL-13 cells by recombinant human IL-4 and
IL-13 was assessed by measuring the levels of SEAP secreted in the supernatant using
QUANTI-Blue .
PRODUCT QUANTITY CAT. CODE
HEK-Blue IL-4/IL-13 Cells 5-7 x 10 6 cells hkb-stat6
HEK-Blue IL-4/IL-13 Kit 1 kit hkb-stat6-kit