Luckily this week we put our usual order of cells on hold, as we needed to analyse the data from the previous immunoprecipitation experiments before making a decision on what to immunoprecipitate with next, but also we had a few samples left in the freezer to use from previous experiments. I have heard that other staff in the lab have had their samples infected with yeast, unlucky and frustrating for them, but a bit of a forewarning to me- don’t brew my own beer or make bread whilst growing up cells.
I’m actually HALF way through my placement now, and I can honestly say I feel like I know Western blot like the back of my hand, and probably immunoprecipitation and other bits and pieces surrounding running gel electrophoresis for Western blotting too.
I learned Densitometry this week, including how to photograph my blots to calculate the relative amounts of protein present in each band after quantifying the blots using actin or ZIP7 as our primary antibody, for normal and immunoprecipitated blots respectively. Also I’ve been using Paint Shop Pro software to enhance and clarify my images for presenting in a final project.
No fun pictures of me doing lab work this week I’m afraid, so I’ve put some examples of the blots I’ve developed (can’t describe in detail as it may need to contribute to a publication), as I know people generally prefer to look at a picture instead of read my rambling on. Not a lot of big news this week, but that usually comes out on a Friday when we’ve had our weekly meeting and tied up loose ends so I’m sure I’ll be updating you on that in my blog next week (which I will publish as normal on late Friday afternoon). Also I will have plenty of photographs in my next blog… I think I’m going to do it like a diary… “A day in the life of me” kind of thing.
Here are some pretty pictures in the mean time for you to ponder over.
Above is some Nitrocellulose paper, which has been stained with Ponceau S red dye to show up the proteins that have been blotted from my gel, to the paper.
And this is a photograph which I developed in the dark room, showing up some bands of an antibody I have used, which I've stained with a Chemiluminescent reagent in order for my antibody to glow in the dark.
Hope you enjoyed this post and are looking forward to my blog next week, which WILL be full of pictures.