The visualization of protein on SDS-PAGE was to be done on the third day so I was excited and slightly nervous as it would signify if I have achieved the aim of the workshop which was Expression of Recombinant Protein. It was on the third day, that I could see the results of the two days of my hard work.
Day 3 of the workshop started with fetching the casted SDS-PAGE gels, supernatant and pellet of lysate which contained the desired protein. The supernatant and pellet were mixed with loading dye and boiled for some time followed by loading the sample on SDS-PAGE. After loading the samples we had to wait for an hour and a half to stain the gel. This one and half hour Anjali ma’am addressed our doubts and gave a short talk on what are the applications of attending such workshops. Now comes the staining step the most crucial step. We performed the two most common staining methods i.e. coomasie blue and silver staining.
Coomasie blue staining was used to stain one gel. This staining showed several proteins on the gel. In the lane the brightest band is the recombinant protein. In every lane there is one bright band which signifies each of us got the recombinant protein.
Silver staining method is quite tricky as we need to stop the development reaction when the bands are moderately visible else it will turn the whole gel brownish black and we won’t be able to notice any bands. But this development reaction is very beautiful to witness, the formation of brown bands in the colourless solution on transparent gel is amazing!
The certificates of the workshop were distributed by Anjali ma’am. Lastly some of the students shared their experience with this we ended our workshop.This workshop was indeed an amazing one and hoping to attend more such workshop and gain more practical knowledge.
- Pooja Shah