Search and Browse
Download
Enter Data
|
Assay Method Information |
|
| | Biological and Stability Data |
| Description: | In vitro assays were conducted with Compounds 1-9 and the commercially available TLR4 agonist MPL. For measurement of biological activity various cells were stimulated with a wide dose range of each compound followed by assessment of either transcriptional activation (HEK hTLR4 NF-κB-SEAP cells) or cytokine production (hMM6 or hPBMCs). Dose-response curves for each compound were started at either 100 μM or 20 μM followed by 5-fold serial dilutions in vehicle (2% glycerin or glycine, IN ) with final concentrations being 1.6 10−8 μM (1.6 fM) or 3.3 10−8 μM (3.3 fM). After incubation for 18-24 h with the dose range of compounds cellular supernatants were harvested for analysis.hTLR4 activation. HEK hTLR4-expressing cells were treated with 100 μM concentration of test compound followed by a 5-fold dilution series. HEK hTLR4-expressing cells also contained an NF-κB driven SEAP reporter and were stimulated with the indicated concentration (FIG. 1A-1C) of the test compound for 18 hours followed by assessment of the cellular supernatant for SEAP by a Quantikine SEAP assay (InvivoGen). The SEAP assay was used to look at secretion of the NF-κB driven alkaline-phosphatase reporter gene in response to TLR4 activation by the compounds and results are interpreted both in terms of potency of the compounds to induce SEAP activation (i.e. potency where a lower EC50 indicates higher potency) and efficacy for receptor activation (i.e. maximal SEAP induction). EC50 values for each compound in HEK hTLR4 cells are shown in Tables 1a and 1b. EC50 values were determined by fitting dose response curves to a non-linear 4-parameter equation. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |
|
Home |
| |
Search |
| |
Deposit |
| |
SiteMap |
| |
About us |
| |
Email us |
| |
Info |
|
©2000 BindingDB. All rights reserved. |
|||||||||||||