Protein Expression in Cells
Quantify fluorescence intensity on a cell-by-cell basis using nuclear and cellular markers
One of the most common assays in cell biology is the protein expression.
This is a very basic tool in cell biology which can be applied for numerous biological assays, such as:
- Dose response assays
- Protein degradation
- Protein accumulation
- Transfection efficiency
and many more.
Quantify fluorescence intensity on a cell-by-cell basis using nuclear and/or cytoplasmic markers.
Cells in each image are automatically identified and their fluorescence intensity is measured.
WiSoft Athena Image analysis software’s fast and labor reducing enables users to rapidly screen a large variety of experimental conditions, as with transfection for example, that uses comparative protein expression as a readout.
A heat-map overlaying the plate layout presents the comparative results for intuitive data visualization.
Enables simultaneous studies of mixed populations.
Protein Expression is suitable for assays such as:
- Cell Health / Autophagy assays
- Protein Localization & translocation
- Subcellular distribution analysis
- Protein degradation/ Accumulation assays (t1/2) / Ubiquitination
- Dose response assays
- Transfection efficiency assay
- Gene therapy assays
- Visualize Response to therapies & drug mechanisms
Protein Expression Athena application applied for detection of cell viability (Autophagy), protein expression and protein phosphorylation Assay in breast cancer research:
Source:
Booth et al., Oncotarget, 26;9(5):6062-6074, Dec 2017
Breast cancer drug neratinib has been found to block the function of the Ras genes which are among the first to be linked to cancer development.
Image analysis showed Neratinib down-regulating the expression of mutant K-RAS and mutant N-RAS.
Neratinib and HDAC inhibitors interact to reduce the expression of mutant N-RAS and mutant K-RAS. Spiky and PANC1 cells that express a mutant N-RAS and a mutant K-RAS, respectively, were treated with vehicle control and neratinib (50 nM). The cells were fixed in place and immunostaining performed with DAPI counter-stain to determine the expression levels and cellular localization of N-RAS and K-RAS at 60× magnification using Hermes HCS system.
“In-Cell Westerns” Assay
A novel approach to detect gross changes in protein expression and protein phosphorylation
Source:
Rationally repurposing Ruxolitinib (Jakafi®) as a solid tumor therapeutic
Tavallai Mehrad, Booth Laurence, Roberts Jane L, Poklepovic Andrew, Dent Paul ; Frontiers in Oncology ,vol.6,2016
Assessments of total protein expression or post translational modifications, such as phosphorylation, can be made via fluorescence intensity measurements. Such “in-cell western” analysis uses unbiased pre-programmed electronic data acquisition, much as has previously been performed for the last 45 years using SDS-PAGE and western immunoblotting.
The table above compares main differences between the conventional western blot assay and protein expression endearments using the Hermes high content imaging system.
One of the most common assays in cell biology is the protein expression.
This is a very basic tool that can be applied for numerous assays, such as dose response assays, protein degradation, Transfection efficiency and many more.
Quantify fluorescence intensity on a cell-by-cell basis using nuclear and/or cytoplasmic markers.
Cells in each image are automatically identified and their fluorescence intensity measured. A heat-map overlaying the plate layout (top) presents the comparative results for intuitive data visualization.
Athena’s fast and labor reducing enables users to rapidly screen a large variety of experimental conditions, as with transfection for example, that uses comparative protein expression as a readout.
Enables simultaneous studies of mixed populations
Protein Expression is suitable for assays such as:
- Cell Health / Autophagy assays
- Protein Localization & translocation
- Subcellular distribution analysis
- Protein degradation/ Accumulation assays (t1/2) / Ubiquitination
- Dose response assays
- Transfection efficiency assay
- Gene therapy assays
- Visualize Response to therapies & drug mechanisms
Source:
Booth et al., Oncotarget, 26;9(5):6062-6074, Dec 2017
Protein Expression Athena application applied for detection of cell viability (Autophagy), protein expression and protein phosphorylation Assay in breast cancer research:
Breast cancer drug neratinib has been found to block the function of the Ras genes which are among the first to be linked to cancer development.
Image analysis showed Neratinib down-regulating the expression of mutant K-RAS and mutant N-RAS.
Neratinib and HDAC inhibitors interact to reduce the expression of mutant N-RAS and mutant K-RAS. Spiky and PANC1 cells that express a mutant N-RAS and a mutant K-RAS, respectively, were treated with vehicle control and neratinib (50 nM). The cells were fixed in place and immunostaining performed with DAPI counter-stain to determine the expression levels and cellular localization of N-RAS and K-RAS at 60× magnification using Hermes HCS system.
Source:
Rationally repurposing Ruxolitinib (Jakafi®) as a solid tumor therapeutic
Tavallai Mehrad, Booth Laurence, Roberts Jane L, Poklepovic Andrew, Dent Paul ; Frontiers in Oncology ,vol.6,2016
“In-Cell Westerns” Assay
Assessments of total protein expression or post translational modifications, such as phosphorylation, can be made via fluorescence intensity measurements. Such “in-cell western” analysis uses unbiased pre-programmed electronic data acquisition, much as has previously been performed for the last 45 years using SDS-PAGE and western immunoblotting.
App note-A novel approach to detect gross changes in protein expression and protein phosphorylation
The table above compares main differences between the conventional western blot assay and protein expression endearments using the Hermes high content imaging system.