Analysis of our Western Blots was performed in approximately 30 seconds using the fully automated process. The program is easy to navigate; analyses were performed with confidence by myself and by graduate students after a five minute demonstration
Dr Roz Jenkins
Principal Experimental Officer, Molecular and Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, UK
Accurate and reliable measurement of protein expression with Western blots requires optimisation and control of multiple steps. CLIQS offers you two common normalisation methods to overcome any technical variation you introduce from sample handling, running 1D SDS PAGE gels and imaging: 1. House-keeping proteins (HKP) 2. Total protein normalisation (TPN) The graph shows an increase in signal achieved by normalising the target protein compared to the results with no normalisation of the target protein signal.
In the example on the left a house-keeping protein GAPDH (green) is measured in one channel and used to normalise a dilution series for a protein of interest in a second channel (red). The graph shows an increase in signal achieved by normalising the target protein compared to the results with no normalisation of the target protein signal.
Western blots are a common way to validate published proteomics discoveries. In this case CLIQS was applied to interpret subtle, rapid changes in post-translational modifications (PTMs) measured by iTRAQ with limited sample and time while accounting for inter-experimental variation. As part of a case study the benefits CLIQS gave were:
CLIQS is a simple affordable software to analyse separation and quantification of your DNA, RNA and protein samples including multiplex western blots. You can import a wide range of image file formats, see the results on the raw image and record the measurements in exportable data tables as you apply each of the steps here.