scaricalo in formato PDF - labogen srl

HEK-Blue IL-6 Cells - IL6 Reporter Cells
➥ Monitor the JAK/STAT-3 pathway
➥ Detection of human IL-6
Background
The proinflammatory cytokine IL-6 is one of the most important
mediators of fever and of the acute phase response. IL-6 exerts its
action by first binding to the IL-6R. The complex of IL-6 and IL-6R
associates with the signal-transducing membrane protein gp130, thereby
inducing its dimerization. This leads to the activation by phosphorylation
of the tyrosine kinases of the Janus family (JAK1, JAK2, and Tyk2).
Activated JAKs induce the dimerization and translocation to the nucleus
of STAT3 where it binds enhancer elements of IL-6-inducible genes.
Description
HEK-Blue IL-6 cells allow the detection of bioactive IL-6 by
monitoring the activation of the STAT-3 pathway. These cells were
generated by stable transfection of HEK293 cells with the human
IL-6R gene. They were further transfected with a SEAP reporter gene
under the control of the IFN-b minimal promoter fused to four STAT3
binding sites. Upon IL-6 stimulation, HEK-Blue IL-6 cells trigger the
activation of STAT3 and the subsequent secretion of SEAP. SEAP can
be readily monitored when using the SEAP detection medium
QUANTI-Blue .
Stimulation of HEK-Blue IL-6 cells with recombinant human IL-6 can be
blocked by anti-hIL-6-IgA, a neutralizing monoclonal antibody of the IgA
isotype (see page 104).
• Detection range for IL-6: 0.5 - 50 ng/ml
HEK-Blue /IL-6 cells are resistant to both hygromycin and Zeocin .
Contents and Storage
HEK-Blue IL-6 cells are grown in standard DMEM medium, 2 mM
L-glutamine, 10% FBS supplemented with 200 µg/ml HygroGold
(ultrapure hygromycin) and 100 µg/ml Zeocin . Cells are provided frozen
in a cryotube containing 5-7 x 10 6 cells and supplied with 100 µl of
HygroGold at 100 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-6 cells are also available in a kit. See below:
HEK-Blue IL-6 Kit
• HEK-Blue IL-6 Cells 5-7 x 10 6 cells
• Anti-hIL-6-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: JAK/STAT3 signaling pathway.
Figure 2: Stimulation of HEK-Blue IL-6 cells by recombinant human IL-6 was assessed
by measuring the levels of SEAP using QUANTI-Blue .
PRODUCT QUANTITY CAT. CODE
HEK-Blue IL-6 Cells 5-7 x 106 cells hkb-il6
HEK-Blue IL-6 Kit 1 kit hkb-il6-kit
109
CYTOKINE SIGNALING 4