scaricalo in formato PDF - labogen srl

INNATE IMMUNITY 3
90
HEK-Blue LPS Detection Kit
Lipopolysaccharide (LPS), also known as endotoxin, the major cell wall component of Gram-negative bacteria, induces the activation of NF-kB
and the production of proinflammatory cytokines. In vivo, this response can cause fever, septic shock and eventually death of the animal. In vitro,
it can introduce a bias in experiments involving cells sensitive to low levels of LPS such as monocytes. In addition, repeated passages of cell
lines in a medium containing LPS might render these cells unresponsive to further stimulation by LPS. This desensitization, termed LPS tolerance,
does not only affect inflammatory responses but also other essential functions including antigen presentation by monocytes. Thus, monitoring
the presence of LPS in biological reagents is crucial. InvivoGen provides the HEK-Blue LPS Detection Kit, a simple, rapid and reliable system
to detect the presence of endotoxins in your samples.
➥ Detection range: 1.5 EU/ml in the sample
Description
The HEK-Blue LPS Detection Kit is based on the ability of TLR4 to
recognize structurally different LPS from gram-negative bacteria and in
particular lipid A, their toxic moiety. Proprietary cells engineered to
become extremely sensitive to LPS, called HEK-Blue -4 cells, are the main
feature of this endotoxin detection kit. The presence of very low
concentrations of LPS, starting as low as 0.03 ng/ml, are detected by the
HEK-Blue -4 cells leading to the activation of NF-kB. Using HEK-Blue
Detection, a specific detection medium, NF-kB activation can be observed
with the naked eye or quantified by reading the OD at 650 nm.
Key Features
HEK-Blue -4 cells, the endotoxin sensor cells, are engineered HEK293
cells. These cells stably express TLR4, MD2, CD14 and multiple genes from
the TLR4 pathway. They coexpress an optimized SEAP reporter gene,
placed under the control of a promoter inducible by the transcription
factors NF-kB and AP-1.
HEK-Blue Selection is a solution that combines several selective
antibiotics. These antibiotics guarantee the persistent expression of the
various transgenes introduced in HEK-Blue -4 cells. Furthermore,
Normocin is included in the kit to protect HEK-Blue -4 cells from any
potential microbial contamination, whether caused by mycoplasmas,
bacteria or fungi (see page 11).
HEK-Blue Detection is a medium specifically designed for the detection
of SEAP. It contains a color substrate that produces a purple/blue color
following its hydrolysis by SEAP (see page 15).
Contents and Storage
The HEK-Blue LPS Detection Kit is composed of the following
components:
- 1 vial of HEK-Blue -4 cells (3-5x 10 6 cells)
- 4 tubes of 250X HEK-Blue Selection (2 ml)
- 4 tubes of 500X Normocin (1 ml)
- 2 pouches of HEK-Blue Detection (50 ml each)
- 1 tube of E. coli K12 LPS, LPS-EK, (100 μg) as a positive control
- 1 tube of endotoxin-free water (1.5 ml) as a negative control
Buy the HEK-Blue LPS Detection Kit once then reorder only the
reagents to perform further assays.
www.invivogen.com/innate-immunity-pamps
HEK-Blue LPS Detection Kit Procedure
PRODUCT QTY CAT. CODE
HEK-Blue LPS Detection Kit 1 kit rep-lps
HEK-Blue Selection 5 x 2 ml hb-sel
Normocin 10 x 1 ml ant-nr-1
HEK-Blue Detection 2 pouches hb-det1
LPS-EK (control) 5 mg tlrl-eklps