Gene Researcher at the Taubert Lab: Day 3

18 03 2012

Today, I learned about Western blot also known as the immuno blot. Just as PCR detects and amplify specific DNA sequences, the western blot detects specific proteins in a sample. I once again used gel electrophoresis to separate proteins by 3-D structure or denatured proteins by the length of the polypeptide. This time I used SDS- Page as in most proteins, the binding of SDS to the polypeptide chain gives an even distribution of charge per unit. I also heated the sample to 60 degrees Celsius to promote protein denaturation and thus, help SDS bind. A tracking dye was added to the protein solution so we could track the progress of the protein solution through the gel during the run. Afterwards, the proteins are transferred to a membrane where they are searched using antibodies specific to the target protein. Since it was my second time making gel electrophoresis, I found easy to pipette the solution into the holes.

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Although reading about Western blotting does not take that long, the actual process actually took the whole day. And in the end, only one of the two samples worked. I was a bit disappointed, but I guess that is what happens in science: experiments don’t always turn out the way you want them to turn out.

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– Alice Yip

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2 responses

19 03 2012
megannantel

Wow the whole day? I guess scientists have to be pretty positive and look at the bright side… ! What can you use the separated proteins for?

28 03 2012
michelle

One of two samples worked? That’s the “glamour” of doing science for real! That is, it’s a lot harder work than it looks like on TV. 🙂 I remember days during my graduate degree where I was further behind at the end of the day than at the beginning. At beginning of the day I had a sample that I’d spent the entire previous day preparing and was ready to test. At the end of the day….well, something had gone wrong, the sample was damaged, and I had to start making a new one. It’s just the nature of experimental science. You have to be persistent.

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