scaricalo in formato PDF - labogen srl

INNATE IMMUNITY 3
HEK-Blue TLR & HEK-Blue NOD Cells
InvivoGen introduces HEK-Blue TLR and HEK-Blue NOD cells, a collection of engineered cell lines designed to provide a rapid, sensitive and reliable method
to screen and validate TLR and NOD agonists or antagonists. They express an NF-kB-inducible secreted embryonic alkaline phosphatase (SEAP) reporter gene
that can be conveniently monitored using the SEAP detection media QUANTI-Blue or HEK-Blue Detection.
HEK-Blue TLR cells
HEK-Blue TLR cells are engineered HEK293 cells that stably co-express a
human or murine TLR gene and an NF-kB-inducible SEAP (secreted
embryonic alkaline phosphatase) reporter gene. To increase the sensitivity
to their cognate agonists, HEK-Blue TLR2 and HEK-Blue TLR4 cells were
further transfected with the co-receptors CD14 and MD2/CD14,
respectively.
HEK-Blue TLR cells are resistant to the selective antibiotics blasticidin
and Zeocin . HEK-Blue TLR2 and HEK-Blue TLR4 cells are additionally
resistant to hygromycin.
HEK-Blue NOD cells
HEK-Blue NOD cells are engineered HEK293 cells that stably co-express
the human and murine NOD1 or NOD2 gene and an NF-kB-inducible SEAP
reporter gene.
HEK-Blue NOD cells are resistant to blasticidin and Zeocin .
HEK-Blue Null cells
HEK-Blue Null cells are the parental cell lines used to generate
HEK-Blue TLR and HEK-Blue NOD cells.
• HEK-Blue Null1 cells express the SEAP reporter gene under the
control of the IFN-b minimal promoter fused to five NF-kB binding
sites.This cell line is the parental cell line of HEK-Blue hTLR2, hTLR3,
hTLR5, hTLR8, h/mTLR9 and h/mNOD1 cells.
• HEK-Blue Null1-k cells express the same reporter system than HEK-
Blue Null1 cells but are slightly different genotypically. This cell line is the
parental cell line of HEK-Blue mTLR3 and hTLR7 cells.
• HEK-Blue Null1-v cells express the same reporter system than HEK-
Blue Null1 cells but are slightly different genotypically. This cell line is the
parental cell line of HEK-Blue mTLR4 and mTLR8 cells.
• HEK-Blue Null2 cells express the SEAP reporter gene under the
control of the IL-12 p40 minimal promoter fused to five NF-kB binding
sites.This cell line is the parental cell line of HEK-Blue mTLR2, hTLR4
and h/mNOD2 cells.
• HEK-Blue Null2-k cells express the same reporter system than HEK-
Blue Null2 cells but are slightly different genotypically. This cell line is the
parental cell line of HEK-Blue mTLR5 and mTLR7 cells.
HEK-Blue Null cells are resistant to Zeocin .
Principle
Recognition of a TLR or NOD agonist by its cognate receptor triggers a
signaling cascade leading to the activation of NF-kB and the production
of SEAP (figure 1). SEAP levels can be determined spectrophotometrically
using HEK-Blue Detection or QUANTI-Blue , both are SEAP detection
media that turn purple/blue in the presence of alkaline phosphatase.
Quality Control
Expression of exogenous TLRs, NOD,1/2, CD14 and/or MD2 was
confirmed by RT-PCR. The functionality of each cell line is validated through
stimulation assays performed with a selection of TLR and NOD agonists.
Note: HEK-Blue TLR, HEK-Blue NOD and HEK-Blue Null cells
express endogenous levels of TLR3, TLR5 and NOD1.
66
PRODUCT
www.invivogen.com/reporter-cells
TLR- and NOD-induced NF-kB signaling pathway
CAT. CODE
(human)
CAT. CODE
(mouse)
HEK-Blue MD2-CD14 Cells hkb-hmdcd NEW -
HEK-Blue TLR2 Cells hkb-htlr2 hkb-mtlr2 NEW
HEK-Blue TLR3 Cells hkb-htlr3 hkb-mtlr3 NEW
HEK-Blue TLR4 Cells hkb-htlr4 hkb-mtlr4 NEW
HEK-Blue TLR5 Cells hkb-htlr5 hkb-mtlr5 NEW
HEK-Blue TLR7 Cells hkb-htlr7 hkb-mtlr7 NEW
HEK-Blue TLR8 Cells hkb-htlr8 hkb-mtlr8 NEW
HEK-Blue TLR9 Cells hkb-htlr9 hkb-mtlr9 NEW
HEK-Blue Null1 Cells hkb-nul11 -
HEK-Blue Null1-k Cells hkb-nul11k -
HEK-Blue Null1-v Cells hkb-nul11v -
HEK-Blue Null2 Cells hkb-nul12 -
HEK-Blue HEK-Blue
Null2-k Cells hkb-nul12k -
NOD1 Cells hkb-hnodl hkb-mnodl NEW
HEK-Blue HEK-Blue
NOD2 Cells hkb-hnod2 hkb-mnod2 NEW
TLR Cells
HEK-Blue NOD Cells
HEK-Blue Null Cells
Contents and Storage
HEK-Blue TLR, HEK-Blue NOD and HEK-Blue Null cells are grown in
DMEM medium, 2mM L-glutamine, 10% FBS and supplemented with
100 μg/ml Zeocin , 30 μg/ml blasticidin and/or 200 μg/ml HygroGold
(ultra-pure hygromycin) depending on the cell line. Cells are provided frozen
in a cryotube containing 5-7 x 10 6 cells and supplied with the corresponding
selective antibiotic(s), 1 ml Normocin (50 mg/ml) and 1 pouch of
QUANTI-Blue . Cells are shipped on dry ice.