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HEK-Blue IL-1b Cells - IL-1b Reporter Cells
➥ Monitor the IL-1b-induced NF-kB and AP-1 pathways
➥ Detection of human and murine IL-1b(a)
Background
Interleukin-1b (IL-1b) is an important mediator of the inflammatory response
to infection and injury. IL-1b binds to the type 1 IL-1 receptor (IL-1R) which
requires the IL-1 receptor accessory protein (IL-1RAcP) to transduce a
signal. IL-1b signaling is mediated by MyD88, IRAK-1/2 and TRAF-6 leading
to the activation of NF-kB and AP-1 (figure 1).
Description
HEK-Blue IL-1b cells allow to detect bioactive IL-1b by monitoring the
activation of the NF-kB and AP-1 pathways. They derive from HEK-Blue
TNF-a/IL-1b cells in which the TNF-a response has been blocked.
Therefore, HEK-Blue IL-1b cells respond specifically to IL-1b. They express
a SEAP reporter gene under the control of the IFN-b minimal promoter
fused to five NF-kB and five AP-1 binding sites.
Binding of IL-1b to its receptor on the surface of HEK-Blue IL-1b cells
triggers the IL1-R signaling pathway leading to the activation of NF-kB/
AP-1 and the subsequent production of SEAP. The levels of SEAP in the
supernatant can be easily monitored using QUANTI-Blue .
HEK-Blue IL-1b cells respond to IL-1b and also IL-1a (data not shown)
but do not respond to TNF-a (figure 2).
Stimulation of HEK-Blue IL-1b cells with recombinant human IL-1b can
be blocked by the neutralizing monoclonal anti-hIL-1b-IgA antibody (see
page 104),
• Detection range for human IL-1b: 100 pg - 100 ng/ml
• Detection range for murine IL-1b: 10 ng - 1 μg/ml
HEK-Blue IL-1b cells are resistant to Zeocin and hygromycin B.
Contents and Storage
HEK-Blue IL-1b cells are grown in DMEM medium, 2 mM L-glutamine, 10%
FBS supplemented with 100 μg/ml Zeocin and 200 μg/ml HygroGold (ultrapure Hygromycin). Each vial contains 5-7 x 10 6
cells and is supplied
with 10 mg Zeocin , 10 mg HygroGold , 50 mg Normocin and 1 pouch
of QUANTI-Blue Cells are shipped on dry ice.
HEK-Blue IL-1b cells are also available in a kit. See below:
HEK-Blue IL-1b Kit
• HEK-Blue IL-1b Cells 5-7 x 10 6 cells
• Anti-hIL-1b-IgA antibody 100 μg
• QUANTI-Blue 1 pouch (100 ml)
• Normocin 1 ml (50 mg/ml)
• HygroGold 100 μl (100 mg/ml)
• Zeocin 100 μl (100 mg/ml)
www.invivogen.com/cytokine-sensor-cells
Figure 1: IL-1b-induced NF-kB signaling pathway.
Figure 2: Stimulation of HEK-Blue IL-1b cells by human and murine IL-1b and
human TNF-a was assessed by measuring the levels of SEAP using QUANTI-Blue .
PRODUCT QUANTITY CAT. CODE
HEK-Blue IL-1b Cells 5-7 x 10 6 cells hkb-il1b
HEK-Blue IL-1b Kit 1 kit hkb-il1b-kit
111
CYTOKINE SIGNALING 4