Dear Protein, I Hate You

I would just like to point out how frustrating it is to come into the lab at 4:30am, all set for yet another absurdly long day of CD, have everything else set up, the small molecule all properly diluted... and then, on the very last step of a long, drawn out, overnight stepwise dialysis....

your protein precipitates.

The protein precipitation doesn't happen in the overnight step, so you can show up first thing in the lab at 4:30am, pull out the cuvette to switch it to the new buffer, see the precipitate, acknowledge that the experiment isn't going to happen, say screw it, and go back to bed. It doesn't happen at the next step at 5:30am either, when you will still justify walking back home and crawling into bed. No, no. It waits until 7:00. After you've already washed out the appropriate pathlength cell with SDS and NaOH to make sure it's all super clean and shiny and ready to go. And you've prepared all your standards, and prepped the small molecule in EtOH (no DMSO for CD experiments!) and then diluted it down in buffer to appropriate concentrations, and have everything on ice and all set, and all you have to do is add the protein once it's out of dialysis and you measure the protein concentration. And 7:00 rolls around and you go to pull out the dialysis cuvette and... wait, what's this? What are all these little white specks floating around that weren't there an hour and a half ago? Why, it's my protein, which decided to crash out just now, even though I've done this same dialysis procedure multiple times before without problems.

And thus, tomorrow, I get to try this all over again. Things like this -- the fact that I work with a protein that has to be purified fresh every 4 days, which requires a somewhat complicated dialysis procedure (which apparently no longer works), which cannot be concentrated AT ALL, which hates to be refolded -- this is why it is going to take me approximately 12 years to complete my Ph.D.

Lab Plague, Part II

Today, my lab took off for the annual lab hike at Old Rag in the Shenendoah Valley. It's a gorgeous hike, and today is a lovely fall day, so I'm sure the lab is quite enjoying themselves. I, however, am not in attendance.

For starters, I probably would have opted out of going this year to begin with, because there's a 1.5ish mile long rock scramble, and my back has been bothering me lately. And considering that it seems like I'll likely be in physical therapy for the rest of my life, and I really aggravated my back doing some hiking and climbing on my honeymoon, doing a 7 mile hike/rock scramble while it's already bothering me doesn't seem like a great idea.

But then there's the issue of being sick. See, the Lab Plague has moved south, and settled into my chest. You know, the deep, phlegm-in-the-lungs coughing, hacking, and wheezing that makes it sound like you're going to drop dead any moment. The constant feeling like you're going to drown in your own mucus, because a giant glob is stuck in the back of your throat, and you're neither able to cough it up, nor swallow it down, and it's just stuck there, in the way.

And yet, despite the fact that I feel (and look) like complete shit, my PI spent the morning yesterday convincing me that I should go on this hike. That I would be totally find. Until, partway through me telling him that there was no way I was up to it, I started coughing. And coughing. And couldn't stop. And slowly, my PI backed out of the lab, index fingers crossed pointed towards me, warding off my germs, and told me to get the hell out of the lab, because when I drop dead, he didn't want it to be in the laboratory (liability issues, I guess).

And so, this morning, I went into lab early to take care of what I needed to do, and then promptly left (because who is going to know that I took the rest of the day off when everyone else is off hiking?), came home, and have spent the rest of the afternoon sprawled out on the couch with a piping-hot mug of tea and my latest knitting project. And a giant lump of mucus in my throat.

Really, and almost lovely afternoon indeed. And my barely-able-to-breath lungs appreciate it much more than the lab hike, even if I am sad to have to miss out on the hiking opportunity once again.

The Plague

I thought I was safe. I thought, for once, I had escaped unscathed. Unharmed. Untouched.
And then, I woke up yesterday morning, unable to breathe. The [name of PI] Lab Plague had officially struck.

This happens, every fall, and then again around February. Someone brings in some unidentifiable flu-like disease. This is no normal cold -- it comes with a fever, and aches, and seriously knocks you down, but it's not the flu -- it's the Lab Plague. And it spreads like wildfire throughout our lab, and only our lab. Pretty much the entire lab, aside from yours truly and the two PIs were out sick the beginning and middle of last week. But since I now sit far, far away from everyone else -- alas, I have an entirely different room number than everyone else -- I thought, perhaps, I would be safe.

Despite my circular dichroism disaster, requiring me to work in the lab for 37.5 consecutive hours, I was feeling fine. Somehow, despite my lack of sleep, my immune system was doing its job! The plague-carriers had taken their sick days off, returned to the lab healthy, and I was still feeling fine. I made it through the weekend completely unsick, and thought I was in the clear.

And then, on Monday morning, I woke up. With a stuffy nose. I tried to convince myself it was just allergies. It would go away. And by 1pm, I was completely unable to breathe, and had developed a fever. By 3pm, I was back home, on the couch, with a headache. And by 9pm, I had been kicked out of my own bed, by a husband, fearful of the plague, forced to sleep on the couch, dreaming of upgrading to a two-bedroom condo, complete with a guest room.

Today, I have not even managed to change out of my pajamas, or even move off the couch, except to make myself a cup of tea (with my brand-new, kiwi green, Le Creuset, whistling teakettle, a wedding present). The floor around me is covered with dozens of used tissues, I'm drinking boiling hot tea, wrapped in a fleece blanket, sniffling, unable to breathe -- and the most bizarre part of it all? Washington, D.C. is in the middle of a heat wave. I have our standard winter Lab Plague, and it is 86 degrees outside. If it didn't require so much energy, I'd go put on shorts, and lie on our patio in the lounge chair and try to bake the plague out of me.

But instead, I used my sick day for some blog maintenance. Finally updated the header to reflect the fact that I am, indeed, now a fourth year grad student, and updated my blog links. If I forgot you -- my apologies -- I'm clearly not thinking well right now. Please, drop me a comment or an email if you belong on my blogroll, and I'll add you ASAP!

But despite the fact that I feel like utter shit, and have zero interest in consuming anything other than the pounds of post-nasal drip falling down the back of my throat, my charming husband just asked me what would be for dinner tonight. Because although the man may be capable of saving lives as a doctor, he is utterly unable to survive on his own or even make grilled cheese. Which means that even though I clearly won't be eating, he feels it is my responsibility as his wife to still cook him dinner. Apparently that whole "in sickness and in health" thing means not that he will take care of me when I am sick, but that regardless of how sick I am... it's still my responsibility to take care of him. Lucky bastard.

#%$*!@$*&!!!!!!!!!!!

I should have known that things were going to go horribly awry this morning when I couldn't even get into the damn building. At 5am, all entrances to the building were blocked off, because they were waxing the floors. There are three different ways to get into this building (four if you count the loading dock, but it wasn't open at 5am), and some genius decided that it was acceptable to wax all the entrances AT THE SAME DAMN TIME. And so, when I attempted to make my way into the building to get to the lab, I was greeted with "Sweetheart, there's wax on that floor, you can't be walking over." I shot a nasty look (don't call me sweetheart), and trudged around to another entrance, where the scene replayed itself.... and then a third time. What the hell?! I voiced my displeasure with the situation, and was told that people are not supposed to be entering the building at 5am. HELLO, I have an ID card that gives me 24 hour access to my laboratory... AS IN, I AM ALLOWED IN THE BUILDING AT 5AM.

So after a very disgruntled hour of sitting in an outside hallway waiting to cross the threshold into my own building, I finally got a late start to my research to what was already going to be a very long day, indeed. I'm chugging along, and went to confirm that I was able to use the CD spectropolimiter. Mind you, I sent an email last Wednesday stating that I wanted to use it on Tuesday and Wednesday, and was that acceptable. I CC'd my PI, and the head of the biochemistry department on this email, and sent it to the post-doc who I go through to use the CD machine. No response, as per usual, but, in the past, if there is a conflict, he'll generally call to say so. I've learned to take no response to mean no conflict. To make sure, I walked over on Friday, but he wasn't around, so I left a post-it note on his desk asking him to please contact me regarding using the machine.

Monday, I went ahead with my protein purification. To get all 3 proteins purified, run on a gel, Coomassie stained, and the dialysis started, is roughly an 18 hour process. They all have to be done back-to-back-to-back, because these proteins cannot be frozen, and last only 2-3 days before degrading. I stopped by again on Monday to confirm the CD, but he had stepped out, but I noticed the post-it note was moved, so obviously he had received it.

I went over this morning to get the key for the room that the spectropolimiter is in, and am told "Oh... you can't do CD today... [chemistry PI] is doing a demonstration on it this afternoon]."

FUCK ME.

You couldn't have told me this Wednesday? Or Thursday? Or Friday? Or even yesterday? It took the entire 10 minute walk from the biochemistry building back to my building of deep breathing to not scream like a banshee and break down in tears. Because now I have minimum 12 hours of CD to do, and I can't start until after 5pm. And I got here at 5am. For absolutely no reason now.

I hope my PI realizes how unbelievably lucky he is to have me, because every other person I've told this story to has said that no way would they be staying from 5pm until at least 5am... they would just call it a day and go home. I'm not entirely sure if it's dedication from my end... or being brainwashed from my PI's end. Probably the latter.

Diary of a Doctor's Wife, Episdoe 003

This afternoon, Husband and I did our weekly grocery shopping trip. It's the one thing we do together (almost every) week, since he's the driver of the relationship, and since I pack all our lunches and cook every dinner that we eat, I can't lug home 1-2 weeks worth of groceries the mile long walk by myself. Husband was on a 36 hour overnight call shift last night, so he asked if we could just go grocery shopping immediately afterward, so he could just come home and sleep for the rest of the afternoon.

I was in lab all morning, so he picked me up in the lab to go grocery shopping, and dropped me back off afterward. Having just come off a call night, he was dressed in his scrubs, so in the checkout line, the female cashier made the educated guess and asked if he was a doctor. She then asked if I was his wife, and then said "So then you're a nurse, right?"

As we walked through the parking lot, Husband commented on the fact that just because I have a vagina, I'm automatically stuck in the position of a nurse. Don't get me wrong -- I'm certainly not stating that there is anything wrong with nursing, and my husband is the first to acknowledge that he couldn't function without the nursing staff -- but why couldn't I have also been a doctor? Why, because I am the female, did I have to be the nurse? But then I also wondered, why did I even have to be a nurse at all? Why didn't she think I was a teacher, or some other equally gender stereotypical role -- was it because doctors are only supposed to marry nurses? Or, more specifically, doctors are supposed to leave their first wives to run off with their nurses (I actually have two friends whose mothers were the nurses and their fathers were the doctors in this exact situation).

Of course, after venting his frustration with society and it's gender role's in the parking lot, he has thus decided to enjoy himself at my expense. This evening, every time he wants something, it has been "Nurse, get me a fork." "Nurse, wash my white coat." "Nurse, bring me some Advil." Unfortunately for him, I am not a nurse, I am a biochemist, and I have access to some extremely toxic drugs in my laboratory. Wouldn't it be a shame if some accidentally wound up in his dinner tomorrow? Whoops. (Just kidding. I know he teases in jest.)