| Redistribution®
assay technology relies on the imaging and quantification
of protein translocation. For development of high quality,
robust, and automation friendly Redistribution®
assays, BioImage uses Aequorea victoria GFP and other
fluorescent proteins fused to translocating targets of interest.
Below
is shown a sketch of the typical reporter construct.
Redistribution®
assays are developed as stable cell lines in order to secure high
assay quality and robustness. We have experience with many commonly
used cell lines – both human and rodent – including:
- U2OS
- HeLa
- T24
- MDA 468
- BHK
- CHO
- And others
See
the complete list of validated Redistribution assays here.
Cytoplasm
to Nucleus Translocation
| Target |
Pathway |
Therapeutic
area |
|
p65 NFkB |
TNF
etc. |
Inflammation |
| ERK1 |
MAPK |
Cancer |
| p38 |
Stress |
Inflammation |
| JNK |
cJun |
Cancer |
| b-catenin |
Wnt |
Cancer |
| Nuclear
receptors |
RXR,
LXR, estrogen etc. |
Many |
| Transcription
Factors |
Transcription |
Many |
|
For
a movie of Erk1 translocation, click
here.
Cytoplasm
to Membrane Translocation
For
a movie of Akt translocation, click
here.
| Target |
Pathway |
Therapeutic
area |
| Akt
(PKB) |
PI3
kinase |
Cancer,
Diabetes |
| PKCa |
Phosphoinositide |
Cancer |
| PKCß |
Phosphoinositide |
Diabetic
complications |
| PKCe |
Phosphoinositide |
Cardiovascular |
| Arno |
PI3
kinase |
Many |
| b-Arrestin |
GPCRs |
Many |
|
Membrane
to Cytoplasm Translocation
For
a movie of PLCd translocation, click here.
| Target |
Pathway |
Therapeutic
area |
| PLCd |
Phosphoinositide |
Many |
| MARCKS |
PKC |
Cancer,
Diabetes |
| Membrane
Receptors |
Receptor
Internalization |
Many |
|
Redistribution®
Assay Quantitation
Redistribution® assays are quantitative assays
which can be run in standard microtiter plates and give consistent
results from assay run to assay run and across different high
content reading platforms.
The
assay response is measured by capturing an image from an assay
well, and the response is quantified by performing image analysis.
The process is shown here for the FKHR
assay.
Instrument
Platforms for Redistribution®
Redistribution® assays are independent of the imaging
platform and have been shown to give consistent results on a number
of instruments on the market, including:
-
IN Cell 3000 (GE Healthcare)
-
Opera (Evotec Technologies)
-
ArrayScan (Cellomics)
-
Pathway HT (BD Biosciences)
-
Discovery1 (Molecular Devices)
-
Icyte (CompuCyte)
Multiplexing
Redistribution® Assays
Multiplexing refers to technology that enables the simultaneous
measurement of multiple parameters in one test compound well.
Redistribution®
assays can be multiplexed by either expressing two targets in
the same cell line labeled with different colors, or by mixing
different assay cell lines directly in the microtiter plate well.
For
an application note describing multiplexed Redistribution®
assays, click
here.
|
