Poly(I:C) Fluorescein - InvivoGen

TLr3 activation with Poly(I:C) Fluorescein
Poly(I:C) Fluorescein-induced TLR3 activation can be monitored using
TLR3 reporter cell lines, such as HEK-Blue hTLR3 cells.
HEK-Blue -hTLR3 cells are designed for studying the stimulation of hTLR3
by monitoring the activation of NF-kB. These cells were transfected with the
human TLR3 gene and an NF-kB/AP-1-inducible SEAP (secreted alkaline
phosphatase) reporter gene. Levels of SEAP can be easily determined with
QUANTI-Blue (a detection medium that turns purple/blue in the presence of
alkaline phosphatase). Alternatively, HEK-Blue Detection, a cell culture
medium that allows for real-time detection of SEAP can be used. A typical
stimulation curve obtained with QUANTI-Blue is given in figure 2.
- Prepare a HEK-Blue hTLR3 cell suspension (250,000 cells/ml) in
DMEM, 4.5 g/l glucose, 10% (v/v) heat-inactivated fetal bovine serum (30
min at 56ºC), 50 U/ml penicillin, 50 mg/ml streptomycin, 100 mg/ml
Normocin , 2 mM L-glutamine.
- In a 96-well plate, add 180 µl of the HEK-Blue hTLR3 cell suspension
per well.
- Stimulate cells with 10 ng -10 μg/ml Poly(I:C) Fluorescein for 6 to 24 h.
- Determine Poly(I:C) Fluorescein stimulation on TLR3 by assessing
reporter gene expression using QUANTI-Blue or HEK-Blue detection
(see related products).
rELATED PrODuCTS
Product
RAW-Blue
HEK-Blue hTLR3
QUANTI-Blue
HEK-Blue Detection
Poly(I:C) Rhodamine
Poly(I:C) (HMW)
Poly(A:U)
Catalog Code
raw-sp
hkb-htlr3
rep-qb1
hb-det1
tlrl-picr
tlrl-pic
tlrl-pau
Figure 2. HEK-Blue hTLR3 cells were stimulated with increasing concentrations of
Poly(I:C) and Poly(I:C) Fluorescein (50,000 cells/well of a 96-well plate). After
an 18h incubation, NF-kB-induced SEAP activity was assessed using QUANTI-Blue .
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