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Module I: An Overview of Gel-based Proteomics


An Overview of Gel Based Proteomics


Proteomics is the systemic study of entire set of proteins expressed by the genome of an organism under specific conditions in a high-throughput format. In gel-based proteomics the complex proteome of any organism is being separated using poly acrylamide gels. Gel-based proteomics includes classical 1D, native PAGE, and 2D electrophoresis (1-DE and 2-DE), as well as 2D -differencegel electrophoresis (2D-DIGE). 2-DE is a versatile tool for the separation and visualization of complex protein mixtures. It is a protein profiling technique that carries out separation using isoelectric focussing (IEF) in the first dimension, which separates proteins based on their isoelectric points (pI), followed by SDS-PAGE in the second, orthogonal dimension, which leads to the further separation of the proteins based on their relative molecular mass.





 In this module we have provided the technological details of 2-DE for different types of biological samples including human serum in light of disease biomarker detection, bacterial proteome for elucidation of drug action and plant proteome to analyse the stress responses. Technical details for each of the extraction and sample processing procedure have been depicted with the help of videos and animations and their theoretical backgrounds along with the simulations, quizzes, assignments and downloadable materials. You may give any relevant feedback at the end of every experiment.





In order to clearly understand each experiment and make best use of the contents provided, we suggest you to proceed as per the following steps. 


  1. Initially start with the theory section, recall technical knowledge over each step, go through the manual and define an overall design, workflow, to conduct a 2DE experiment.

  2. In protocol section learn the minute integrity required to perform the experiment by going through the standardized protocol defined for each of the steps.

  3. Practice; operate through each step of the experiment in simulator section to visualize the entire process.

  4. Go through the video files to get real lab experiences of performing experiments associated with gel-based proteomics.

  5. In the download sections, course contents are provided for the users to export files, for analyses and in case to perform assignments.  Download the raw gel images provided and perform data analysis using the software. To asses yourself compare your results with the results obtained from our analysis for the same gels.

  6. Now, user should be sufficiently equipped to face a Quiz for assessment. So answer the questions provided in the quiz and complete the assignment. (Go back to any modules that require better understanding based on your performance!)

  7. Students are strongly advised to go through the references provided in the “Reference” section to further advance their knowledge.

  8. Finally provide your feedback on the overall contents which may help us to improve the quality of this e-Learning platform.







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