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CYTOKINE SIGNALING 4
HEK-Blue TNF-a/IL-1b Cells - TNF-a and IL-1b Reporter Cells
➥ Monitor the TNF-a- and IL-1b-induced NF-kB and AP-1 pathways
➥ Detection of human and murine TNF-a and IL-1b(a)
Background
Tumor necrosis factor alpha (TNF-a) and interleukin 1 beta (IL-1b) are
proinflammatory cytokines produced in response to microbial infection.
The common inflammatory responses of TNF-a and IL-1b are mediated
by interactions of these cytokines with distinct receptors: TNF-a binds
TNFR1 and TNFR2, IL-1b binds IL-1RI. Binding of these cytokines to their
receptors induces intracellular signaling. TNF-a signaling involves TRADD,
TRAF2 and RIP, while IL-1b signaling is mediated by MyD88, IRAK-1,
TRAF-6, and Rac1. Both signalings lead to the activation of the
transcription factors NF-kB and AP-1 (figure 1).
Description
HEK-Blue TNF-a/IL-1b cells are designed to detect bioactive TNF-a and
IL-1b (and IL-1a) by monitoring the activation of the NF-kB pathway.
HEK-Blue TNF-a/IL-1b cells derive from HEK293 cells. They
endogenously express the receptors for TNF-a and IL-1b and stably
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.
HEK-Blue TNF-a/IL-1b cells secrete SEAP upon stimulation by human
TNF-a and IL-1b (figure 2). SEAP production is also detected in the
presence of murine TNF-a and IL-1b, and human or murine IL-1a. SEAP
levels can be readily determined using a SEAP detection medium, such
as QUANTI-Blue or HEK-Blue Detection medium.
Stimulation of HEK-Blue TNF-a/IL-1b cells with recombinant human
TNF-a or IL-1b can be blocked by the neutralizing monoclonal antihTNF-a-IgA
or anti-hIL-1b-IgA antibody, respectively (see page 104).
• Detection range for TNF-a: 0.5 ng - 1 μg/ml
• Detection range for IL-1b: 0.2 - 100 ng/ml
HEK-Blue TNF-a/IL-1b cells are resistant to Zeocin .
Contents and Storage
HEK-Blue TNF-a/IL-1b cells are grown in DMEM medium, 2 mM
L-glutamine, 10% FBS supplemented with Zeocin (100 μg/ml). Each vial
contains 5-7 x 10 6 cells and supplied with 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 TNF-a/IL-1b cells are also available in a kit. See below:
110
HEK-Blue TNF-a/IL-1b Kit
• HEK-Blue TNF-a/IL-1b Cells 5-7 x 10 6 cells
• Anti-hTNF-a-IgA antibody 100 μg
• Anti-hIL-1b-IgA antibody 100 μg
• QUANTI-Blue 1 pouch (100 ml)
• Normocin 1 ml (50 mg/ml)
• Zeocin 100 μl (100 mg/ml)
www.invivogen.com/cytokine-sensor-cells
Figure 1: NF-kB/AP-1 signaling pathways induced by TNF-a
and IL-1b.
Figure 2: Stimulation of HEK-Blue TNF-a/IL-1b cells by recombinant human
TNF-a and IL-1b was assessed by measuring the levels of SEAP secreted in the
supernatant using QUANTI-Blue .
PRODUCT QUANTITY CAT. CODE
HEK-Blue TNFa/IL-1b Cells 5-7 x 10 6 cells hkb-tnfil1
HEK-Blue TNF-a/IL-1b Kit 1 kit hkb-tnfil1-kit