2D Gel Image Analysis
2D gel electrophoresis (2-DE) is a key technique for molecular biology. With technology improvements 2-DE has become accessible, affordable and easy to implement. The images you capture from 2-DE gels are quantitative maps of intact proteins that allow you to characterise samples by:
- Relative protein expression changes based on experimental changes
- 2D Western blotting patterns compared to 2-DE protein gels
- pI and molecular weight estimations
- Protein-protein interactions
- De novo sequencing and identification for organisms with incomplete genome sequences
- Post-translational modifications and protein isoforms
Fast, easy to use and reproducible differential protein expression analysis. It is simple to install and apply to support your research in:
- Biomarker Discovery
- Cancer Research
- Food Research
- Evolutionary Biology
- Population Studies
- Environmental Toxicology
- Agricultural Research
- Molecular Biology
Easy-to-use and novel software specifically for analysing biologic product characterisation, process development/optimisation and HCP antibody assay & impurity profiling. Image alignment allows the benefits of a DIGE-like approach without the additional cost or expertise required to use this technique.
Which 2D image analysis software do I need?
|Experiment Type||Differential protein expression analysis of two or more groups of replicate 2D gels||Comparing gels and Western blots to allow spot counting, measuring simple volume changes and reporting relative coverage|
|Image QC/import||Checks images before use for good quality. Allows some image editing||Checks images before use for good quality. Allows image editing including scaling for different size image capture. Useful if Western blots imaged using a different system|
Pixel level alignment to remove positional variation between images.
|Spot Detection||Spots detected in same positions on all gels||A single spot map is created to identify the position of all spots across all images.|
|Matching of Spots between images||100% matching and no missing values. Very good for statistical analysis of differences across groups||The same spot map is applied to all images where spots are categorised as present or absent to allow calculation of percentage coverage.|
|Reporting||Spots are reported using p-values and we are looking for spots that vary the most between experimental groups. Multivariate statistical analysis with no missing values due to alignment and detection approach||Report the percentage coverage and number of spots present for each image compared to a base image. 1 vs 1 image comparisons to identify common and unique spots and comparisons of an area of the spot map are possible. All results and parameters are easily exported from the software.|
- Reducing time and subjectivity of your 2-DE image analysis
- Analysing 2D gels with any type of staining including DIGE and secondary protein stains.
- Increasing sensitivity for measuring low level expression changes in your experiment
- Eliminate missing values in your data for increased statistical power
- Visualisation and quantification of your sample preparation, gel running and imaging quality
- Making discoveries that are real, not just the result of biological variation.
- SameSpots applies pixel level alignment proven to generate reproducible results.
- 100% spot matching all of the time, there is no need to do any match editing.
- SameSpots makes gel analysis more efficient, so you can run enough replicates.
- Multivariate statistical tools included as well as univariate measurements and export to R function.
- Accurate comparison of total protein stained 2D gels with 2D Western blots and reporting of relative coverage
- Helps to meet regulatory requirements by providing additional data on control of biologic quality, potency and safety
- An orthogonal approach to 1D gel and Mass spectrometry.
- Process development and optimisation – allowing for better process decisions to be made to avoid additional costs and delays to production
- Product characterisation with any changes to the product (including post-translational modifications) detected
- HCP antibody assay and impurity profiling, essential for generating process and cell-line specific antibodies
- SpotMap is a single separate analysis tool, not a work around within existing differential expression analysis software.
- Overcomes the problems of using other software packages, such as:
- Experiment set-up limitations
- Trying to compare different spot outlines between blots and gels
- Compatible with file formats from all the common image capture devices.
- Use DIGE, flourometric, chemiluminescent or colorimetric stains in the same simple workflow.
- Checks images quality before analysis with image editing including scaling for different size image capture.
- Compatible with latest Windows operating systems
We’ve been looking at different solutions for three months now and from what we just saw [demonstrated] this is already the bestMajor pharmaceutical company following a first demo of SpotMap