Just a reminder, my name is Colton and I’m visiting the
Buccella Laboratory at New York University. The lab is part of the chemistry
department. More information about the lab can be found at http://www.nyu.edu/fas/dept/chemistry/buccellagroup/
.
Monday morning started off early as usual. As I waited for
everyone to arrive, I ate my breakfast overlooking Washington Square Park. It
is such a beautiful view, but It isn’t just the view that is astounding; it’s a
combination of NYU, their facilities and more importantly the friendly,
intelligent people I’ve been able to work with.
Finally when Sarina arrived,(the graduate student I’ve been
working with), I found out that I would be continuing my mini-project. My
mini-project is fluorescent tests on 3 different compounds in various solvents,
that I mentioned in “Fluorescence; The Crazy World of Wavelengths Week 3”. I
had to finish testing compound 1c, which took up the morning and went into the
afternoon.
The next day I planned to continue the experiment, but there
was a catch. I've come to found that this often happens in research and it is
generally time related. I could technically start, but I couldn't collect data
till the following day. The issue was a matter of equilibration. Compounds 1a
and 1b that I would now be testing had to equilibrate for at least 6 hours in 2
of the solvents I was testing them in, PIPES buffer and water. I prepared these
solutions and all the others, so I would not have to take the time to make them
the following day. After a quick lunch we continued our ATP titrations. A brief
reminder; we are testing a fluorescent compound’s fluorescence when bound to
Magnesium and the effects ATP has on the system. *For a more detailed background
of the experiment please see week 2 and 3 of my blogs. 10 fluorescent scans had to be taken every ½ hour
which meant we were going to be in the lab a while. Combined with our late
start in the afternoon and the amount of time needed between scans, we didn’t
finish the experiment until 8:30pm. It was all worth it in the end, we gathered
the data we needed and found out that we don’t have to wait as long between
scans in future experiments. On the overhand, we were still unsure what exactly
was happening within the system. We believe it could be photo bleaching or slow
equilibration. Photo bleaching is the breaking down of a compound over time as
it is subjected to light (excited). Photo bleaching occurs at different rates
for every compound, and the goal is for a compound to slowly photo bleach,
meaning it can be excited multiple times before truly breaking down.
The next day I was able to continue my mini-experiment on the
UV-Vis & fluorimeter immediately. I have figured out a fairly proficient system
for running my tests, which allowed me to finish them in a few hours.
 |
| Uv-Vis |
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| Fluorimeter |
At group meeting this week, Debbie presented techniques used
in the paper “Labeling Lysine Acetyltransferase Substrates with Engineered Enzymesand Functionalized Cofactor Surrogates”. You can access the article by clicking on its' title above. Although
there was some information I did not have enough biology background to
understand, I found that Debbie did well presenting the information, only
adding to my comprehension. The big picture for me was learning more about the
ways in which biology and chemistry interact.
Next week should be just as exciting as we begin to explore the reasons behind the results of the ATP titrations!
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