SpotMap

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.

SpotMap has been proven to be reproducible and easy to use.

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.

Alignment is a key feature within SpotMap. It removes positional variation between images to allow direct comparisons of spots present on different images to be made.

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

The Proteomics core facility is based at the Rowett Institute but operates as part of the University of Aberdeen. They have recently started using SpotMap in their lab. Read what they think about it here.

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

The reproducibility of coverage results obtained by SpotMap was assessed across three data sets of varying difficulty, by three users of varying expertise.

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.

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Technical specification

Customer testimonials

SpotMap played an integral role in our comparison of proteomes of different cell lines during the process of choosing a proper antigen for antibody development. The ability to overlay gels and assign numbers to each protein spot was especially useful in determining similarities and differences between gels without duplicating identified spots. One feature that I found to be especially useful was the 3D view which allowed for a better visual of spot intensity when determining spots. The table of identified spot for each image also made it easy to identify which spots are unique or common to the different images. With this software, my team was able to make a more informed choice on the best candidate to use for our purposes.

Virginia Liu-Compton

Research Associate 2, ImmunoGen Inc.

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.

 

Rebecca Oettle

PhD Student , University of Cambridge

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