Examples and Evidence
We work closely with our customers from across the globe to ensure they get the best from SpotMap.
To help you to evaluate SpotMap for your projects we can provide you with a demonstration of the software using your own images and fully support you throughout a free trial of the software.
Below is a small sample of reports, posters, application notes and more demonstrating the use of SpotMap.
Example analysis report: SpotMap analysis of a 2D gel vs Western blot
The images used in this analysis were challenging to analyse due to low colour depth, presence of saturation, non-spot features and steaking on the blot image. Issues such as these are commonly experienced in the analysis of 2D gels and Western blots. SpotMap provided results quickly, within an hour, to calculate percentage coverage of the 2D gel by Western blot as 17% following identification of 709 spots.
Technical note: Removal of positional variation using alignment.
In this analysis the use of two different alignment targets were compared. In one analysis the Western blot was aligned to the gel. In the other analysis, both the 2D gel and Western blot were aligned to the stained membrane of the Western blot.
14% coverage of the 2D gel was recorded following both methods of alignment.
Application note: Anti-HCP antibody characterisation using SpotMap
SpotMap was used to characterise two pooled samples of antisera by comparison of Western blots to a 2D gel.
Percentage coverage of each pooled antisera was calculated and spots recognised were compared. Pool 1 antisera had a broader reactivity at 44% coverage, compared to 39% coverage by pool 2. Comparisons of low and high molecular weight regions were also made. Pool 1 had lower coverage in the low molecular weight region.
Case Study: University of Aberdeen, Proteomics
2D-SDS PAGE analysis methods for determining HCP Coverage using process derived anti-HCP polyclonal antibodies
A silver stained 2D gel was compared to a Western blot captured at 3 exposure times of 1.0, 3.2 and 5.5 seconds. Analysis was completed using PDQuest and SpotMap, the workflow of each software program and percentage coverage results were compared. Analysis using PDQuest took approximately 3.5 hours and provided percentage coverage results in a range of 11% across the three exposure times. SpotMap analysis took approximately 1 hour and provided percentage coverage results in a range of 1% across all exposure times.
Inter-user variation of percentage coverage is within a range of 2% using an optimised analysis process
SpotMap software used to minimise inter-user and inter-lab variation of anti-HCP antibody percentage coverage results
Users with a range of experience at two locations analysed two data sets of a 2D gel vs Western blot and the percentage coverage results were compared.
The coefficient of variation for each analysis was within 10%. Key areas to be considered for future analysis and creation of an SOP are also addressed.
Originally presented at WCBP 2016.
I have found the software very easy to use and has been exactly what I needed to compare western blots and pick the spots from my gel, which have now been sent off for mass spec.