HEK-Blue™ LPS Detection Kit
Cell-Based Assay for the Detection of Endotoxins in Biological Samples
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-κB. Using HEK-Blue™ Detection, a specific detection medium, NF-κB activation can be observed with the naked eye or quantified by reading the OD at 650 nm.
Specifications
Requires only basic cell culture knowledge
Hands-on time: < 1 hour. Gives results after overnight incubation
Detects only endotoxins - No false positive
Up to 500 samples can be tested with the kit. To perform further assays, only the reagents need to be reordered
Detection range: 1.5 EU/ml in the sample
Contents
- 1 vial of HEK-Blue™-4 cells (3-5x 106 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
Description
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 repor ter 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 selectiveantibiotics. These antibiotics guarantee the persistent expression of the various transgenes introduced in HEK-Blue™-4 cells. Fur thermore, Normocin™ is included in the kit to protect HEK-Blue™-4 cells from any potential microbial contamination, whether caused by mycoplasmas, bacteria or fungi.
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.
Details
LPS Detection using the HEK-Blue™ LPS Detection Kit
The presence of LPS in various preparations of TLR ligands was determined using the HEK-Blue™ LPS Detection Kits: 1) Monophosphoryl lipid A, 2) LPS-EB (E. coli 0111:B4), 3) LPS-EK (E. coli K12), 4) LPS-SM (S. minnesota), 5) LPSPG (P. gingivalis), 6) ODN 2006, 7) E. coli DNA ef (endotoxin-free), 8) E. coli DNA we (with endotoxin).
Ref.
- Zhang G. and S. Ghosh, 2000. Molecular mechanisms of NF-kappaB activation induced by bacterial lipopolysaccharide through Toll-like receptors. J Endotoxin Res. 6(6): p. 453-7.
- Beutler B., 2003. Science review: key inflammatory and stress pathways in critical illness - the central role of the Toll-like receptors. Crit Care. 7(1): p. 39-46.
- Dobrovolskaia M.A. and S.N. Vogel, 2002. Toll receptors, CD14, and macrophage activation and deactivation by LPS. Microbes Infect. 4(9): p. 903-14.
- Wolk K. et al., 2000.Impaired antigen presentation by human monocytes during endotoxin tolerance. Blood. 96(1): p. 218-23.
- Kariko K. et al., 2004. Small interfering RNAs mediate sequence-independent gene suppression and induce immune activation by signaling through toll-like receptor 3. J Immunol. 172(11):6545-9.
- Darveau RP et al., 2004. Porphyromonas gingivalis lipopolysaccharide contains multiple lipid A species
that functionally interact with both toll-like receptors 2 and 4. Infect Immun. 72(9):5041-51.
ORDERING
HEK-Blue™ LPS Detection Kit
| Cat. Code | rep-lps |
|---|---|
| Unit Size | 1 kit |
| Price | For price or distributor address, please select your country |
