Protein expression can be used to measure the way in which proteins are synthesised, modified and regulated in living cells. The modulation of protein expression by project compounds can provide powerful information to help guide your discovery program. At Sygnature Discovery we have experience in measuring protein expression levels using a variety of assay formats.
Protein Expression Levels
Established techniques at Sygnature for the measurement of protein expression levels include:
- Dot Blot
- Western Blot
- Traditional ELISAs
Dot-blot techniques can be used to visualise and measure protein expression levels from large volume lysate loads, this has particular benefit if the target of interest is only present at low levels within the sample.
- 96-well manifold allows for higher sample throughput than a traditional SDS-PAGE based analysis.
- Advantageous when analysing compounds IC50 values or optimising conditions.
- Enables direct loading from the culture plate.
Western blotting is a routine technique for protein analysis, where the specificity of the labelled antibody-epitope interaction enables a target protein to be identified within the midst of a complex protein mixture.
Western blotting can produce qualitative and semi-quantitative data about the protein of interest, detected using our ChemiDoc™ MP Gel Imaging System.
As well as traditional western blot methods, Sygnature can also employ our Simple WesternTM system for high-throughput and quantitative work.
The Simple WesternTM system offers a more automated and higher-throughput approach to western blotting by utilising the capillary based separation of protein targets by size. The process removes the tradition hands-on gel and blotting steps and replaces these with automated separation and detection steps that enables data analysis after just three hours.
The Jess instrument allows the analysis of 24 samples per run with immunodetection available through one chemiluminescence and two fluorescence (IR and NIR) channels, enabling the multiplex analysis of different protein targets of interest from a single sample. Further to this, protein normalisation can also be conducted without compromising on the available detection channels, measuring total protein content of the sample and allowing easy run-to-run comparison.
Applications include the quantification of target protein phosphorylation and DC50 and Dmax determination for targeted protein degradation assays.