VIDEO

1. Introduction to Gel based Proteomics

2. Serum Protein Extraction

This video depicts the process of blood collection, serum separation, sonication of serum proteins, removal of high abundance proteins and finally, protein precipitation.

3. Protein Quantification

Following protein extraction, the separated proteins must be quantified. Here, we depict the process of quantification of standard and sample proteins using the Bradford assay.

4. First Dimension- Isoelectric Focusing

This video demonstrates the steps involved in isoelectric focusing, which separates proteins in their first dimension based on their isoelectric points. This includes rehydration of the IPG strips and focusing.

5. Second Dimension - SDS PAGE

This video depicts the steps involved in separation of proteins in the second dimension based on their molecular weight. It involves assembly of the gel apparatus, gel casting, equilibration of the IPG strip and electrophoresis.

6. Staining and Destaining

After completion of electrophoresis, the gels must be stained for visualization of the spots. This involves removal of the gel from the 2-DE unit, fixing of protein bands, staining with suitable dye solution and finally, destaining to view the stained protein bands.

7. Data Analysis

This video takes the user through the process of gel data analysis in a stepwise manner using a 2-DE analysis software. This will enable the user to perform analysis independently when provided with raw gel images and suitable software.