It’s been a few weeks since starting our phased return to the labs here at Leeds. I’m very fortunate to be part of the early phase of returning. I’ve been very eager to get back in the lab, I definitely missed the lab during lockdown but I was also pretty nervous about heading back. It’s been over 4 months since I last picked up a pipette and ran a western blot, among many other lab activities. I was apprehensive about all the changes and generally just being around more people. I’d kind of gotten used to my own little bubble of people during lockdown it almost seems weird to venture out of that.
Of course, things were different, there’s about 40 of us as opposed to over 100 people in the building. The 40 of us are staggered over two days throughout the week so there aren’t many people in at any one time. It’s very quiet, almost too quiet. In some ways, it’s super peaceful and you can crack on with what you’re doing but the temptation to chat to people while passing in the corridor is quite high but unfortunately, that’s not allowed. Our uni is following a strict 2-metre rule which, to be honest, I’m quite happy about, I know that makes things a little harder in terms of logistics and space but it’s certainly much safer, and I feel much better about it. We also have to wear masks in the lab, which at first I wasn’t too pleased about with being someone who’s hearing impaired but actually, it’s really not that bad and because there’s not much chatting going on it’s really not a problem. Just to be clear I’m not anti-mask under any circumstances but sometimes I just struggle to hear people, that’s all.
Again, I’ve been pretty lucky in the sense that my supervisor has his own little lab which I essentially had to myself in the two days I was in. It was nice, I had the radio on, didn’t have to wear a mask and I wasn’t at risk of being in anyone’s way. But I do have to say I miss my work colleagues!
Typically I rely heavily on blood donations to do any work but unfortunately, we’re unable to take blood at the moment so what can I possibly do in the lab you say? Well, I have to seriously thank past Beth for batching Western Blot samples and creating an inventory. So I’m spending most of my time working through Western Blot samples.
What are Western Blots?
Western Blots are used to detect specific protein molecules among a mixture of proteins within cells or tissue. In my case, the cells I’m using are platelets which are cells of the blood. I’m blotting for particular phosphorylation events within platelets.
SDS-PAGE (electrophoresis) which is where we separate the proteins based on size. The blue stuff is called Laemmli buffer and that’s what we use to lyse (pop) our cells and to make their charge negative. The samples are run across a gel, with proteins migrating towards the positive electrode. Smaller proteins travel further & faster whereas larger proteins travel less & slower. After the gel process, we then transfer the proteins to a membrane that allows us to probe using antibodies for specific proteins.
What are phosphorylation events?
Phosphorylation is a reversible process where enzymes called phosphatases remove phosphate groups from proteins. Phosphorylation is essentially a molecular switch which can modify the activity of a protein. In my case, I’m looking at inhibitory phosphorylation events that aid platelet inhibition.
Like most biology PhD students, I have a love-hate relationship with Western Blots. There are a lot of steps involved and you’re only as good as your antibody as my supervisor so frequently says. At the moment I’m just super happy about being back in the labs!
I have to say I was definitely pretty rusty, I used the wrong pipette first thing (I used a p200 instead of p20 – which is quite different!). And, my blots didn’t turn out very well. I think it’s a combination of being a bit rusty and new imaging software. I’m not being too hard on myself, yes it is a bit annoying as I tend to be the self-proclaimed “blot queen” but let’s be real, it’s been a long time since I last did one so no hard feelings. I’ll try again next week!
I’d love to know if you’re back in the labs yet? If so, what experiments are you doing? Do you like Western Blots? What’s your favourite experiment?
Thanks for reading and I hope you enjoyed this post!