|
| 1. |
Tang, Y., Scheef, E.A., Wang, S., Sorenson, C.M., Marcus, C.B., Jefcoate, C.R. and Sheibani, N.
(2009)
CYP1B1 expression promotes the proangiogenic phenotype of endothelium through decreased intracellular oxidative stress and thrombospondin-2 expression.
Blood
113
,
744–754
.
|
| |
Notes:
The authors tested if CYP1B1 removed cellular oxygenation products that induce oxidative stress and promote the release of antiangiogenic factors. The P450-Glo™ CYP1B1 Assay was used to determine CYP1B1 activity. The presence of glutathione was assessed using either 104 retinal endothelial cells or 50µl of mouse retinal extracts dispensed into each well of a 96-well plate with the GSH-Glo™ Glutathione Assay.
(0004010) |
| |
 |
| |
Products: GSH-Glo™ Glutathione Assay | P450-Glo™ CYP1B1 Assay |
| 2. |
Fan, F. and Wood, K.V.
(2007)
Bioluminescent assays for high-throughput screening
Assay Drug Dev. Technol.
5
,
127–136
.
|
| |
Notes:
The authors of this paper review bioluminescent assay technologies, discussing HTS reporter, cell-based and luciferase biosensor assays. They divide luminescent assays into three basic categories: assays that measure ATP concentration (cell viability and kinase assays), assays that measure changes in luciferase levels (reporter assays, GPCR assays), and assays that measure changes in luciferin levels (protease [including caspase], P450 and MAO assays).
(0003737) |
| |
|
| |
Products: Renilla Luciferase Assay System | BacTiter-Glo™ Microbial Cell Viability Assay | Bright-Glo™ Luciferase Assay System | Calpain-Glo™ Protease Assay | cAMP-Glo™ Assay | Caspase-Glo® 2 Assay | Caspase-Glo® 3/7 Assay | Caspase-Glo® 6 Assay | Caspase-Glo® 8 Assay | Caspase-Glo® 9 Assay | CellTiter-Glo® Luminescent Cell Viability Assay | Dual-Glo® Luciferase Assay System | Dual-Luciferase® Reporter 1000 Assay System | Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | Kinase-Glo® Lumine |
| 3. |
Tsuchiya, Y., Nakajima, M., Takagi, S., Taniya, T., and Yokoi, T.
(2006)
MicroRNA regulates the expression of human cytochrome P450 1B1.
Cancer Res.
66
,
9090-9098
.
|
| |
Notes:
These authors identified a region complementary to the microRNA miR-27b in the 3´ UTR of the cytochrome p450 CYP1B1 mRNA, and showed that miR-27b was involved in regulation of CYP1B1 expression. The 3´ UTR containing the miRNA target site was cloned downstream of the luciferase gene in the pGL3 Promoter Vector and cotransfected into the miR-27b-positive breast cancer cell line MCF-7 and into miR-27b-negative Jurkat cells. Luciferase expression levels from the reporter vector containing the CYP1B1 3´ UTR sequence were reduced in miR-27b-positive cells, but not in the Jurkat cell controls. Delivery of an antisense oligoribonucleotide directed against miR-27b to MCF-7 cells containing the reporter construct resulted in restoration of luciferase activity. The effects of inhibition of miR-27b on protein levels and enzymatic activity of CYP1B1 were then investigated in MCF-7 cells. CYP1B1 protein levels and enzymatic activity increased significantly in cells transfected with the antisense oligo; the enzymatic activity was measured using a p450-Glo™ Assay. The coding region and 3´ UTR of the CYP1B1 gene were also PCR-amplified, subcloned the into the pTargeT™ Mammalian Expression Vector, and transfected into HEK293 cells. The effect of overexpression of miR-27b on protein levels and enzymatic activity of CYP1B1 was then evaluated in these cells.
(0003622) |
| |
|
| |
Products: Dual-Luciferase® Reporter Assay System | Dual-Luciferase® Reporter Assay System 10-Pack | P450-Glo™ CYP1B1 Assay | pGL3-Promoter Vector | pTARGET™ Mammalian Expression Vector System |
| 4. |
Zhang, W., Chen, M., West, D.B. and Purchio, A.F.
(2005)
Visualizing Drug Efficacy In Vivo
Molecular Imaging
4
,
88-90
.
|
| |
Notes:
The authors of this paper present proof-of-concept experiments showing that drug metabolism enzyme activity can be measured in whole animals (in vivo) in real time. Using a mouse that expresses a luciferase transgene at constitutively high levels in the liver, the authors evaluated CYP3A4 and CYP3A7 activity using a CYP3A P450 substrate (proluciferin substrate) that is converted into a luciferase substrate by CYP34 activity. The luciferase substrate produced by the P450 activity is then used by luciferase in a reaction that produces light. An increase in luminescence correlates with an increase in enzyme activity in this assay. The authors conclude that optical imaging of reporter mice will provide a new method for looking at drug actions in whole animals, with the caveat that the solubility of the proluciferin substrate is optimized and toxicity is minimized.
(0003996) |
| |
|
| |
Products: P450-Glo™ CYP3A4 Assay | P450-Glo™ CYP3A7 Assay |
| 5. |
Yueh, M.F., Kawahara, M. and Raucy, J.
(2005)
High volume bioassays to assess Cyp3A4-mediated drug interactions: Induction and inhibition in a single cell line.
Drug Metab. Dispos.
33
,
38–48
.
|
| |
Notes:
A CYP3A4 response element consisting of proximal and distal enhancer sequences with the CYP3A4 promoter was cloned into a pGL3 vector and used in transfection studies with HepG2 cells. The effects of either or both enhancer motifs on luciferase expression were studied in relation to various xenochemical treatments. The researchers also used the P450-Glo™ CYP3A4 Assay to analyze increases in CYP3A4 activities in a stably transfected cell line (DPX-2) and on primary hepatocytes. For these assays, the researchers incubated cells in 96-well or 24-well plates with or without CYP3A4 inhibitors and the P450-Glo™ CYP3A4 substrate. Following a 3 hour incubation, the P450-Glo™ Detection Reagent was added and the luminescence was recorded. In these studies cells were also pretreated with various chemicals including 10µM rifampicin, 1000µM phenobarbital, 100µM dexamethasone, 50µg/ml kava, 50µM methoxychlor, 50µM troglitazone, 100µM omeprazole, 100µM 2-acetylaminofluorene, and 25 µM chrysin.
(0003221) |
| |
|
| |
Products: P450-Glo™ CYP3A4 Assay |
| 6. |
van Duursen, M.B., Sanderson, J.T. and van den Berg, M.
(2005)
Cytochrome P450 1A1 and 1B1 in human blood lymphocytes are not suitable as biomarkers of exposure to dioxin-like compounds: polymorphisms and interindividual variation in expression and inducibility.
Toxicol. Sci.
85
,
703-712
.
|
| |
Notes:
The authors examined interindividual differences in cytochrome P450 CYP1A1 and CYP1B1 expression levels in human lymphocytes before and after exposure to dioxins and dioxin-like compounds. CYP1A1 and 1B1 mRNA levels were assessed with semi-quantitative RT-PCR using the Access RT-PCR System.
(0003432) |
| |
|
| |
Products: Access RT-PCR System |
| 7. |
Rodriguez-Melendez, R., Griffin, J.B. and Zempleni, J.
(2004)
Biotin supplementation increases expression of the cytochrome P450 1B1 gene in Jurkat cells, increasing the occurrence of single-stranded DNA breaks.
J. Nutr.
134
,
2222–2228
.
|
| |
Notes:
Microsomal extracts from Jurkat cells were assayed for cytochrome P450 1B1 (CYP1B1) using the P450-Glo™ CYP1B1 Assay. CYP1B1 activity increased when Jurkat cell cultures were supplemented with 10nM biotin.
(0003132) |
| |
|
| |
Products: P450-Glo™ CYP1B1 Assay |