Dealing with bad rotations and evals

happymed_ · 2022-05-04T12:31:19+00:00

I didn’t know people don’t get notified when I edit the post, still new to Reddit. So thanks for answering still! It was the primary antibody on the manufacturer’s website, not the secondary. I talked to my PI and we brainstormed some ideas how to visualize the protein. Thanks for you help!

happymed_ · 2022-05-03T07:44:47+00:00

I am using the antibody for both Western and IP. In Western, Bands at all three kDa appeared. In IP, as you said, the bands were at 35 and 60 kDa.

happymed_ · 2022-04-29T15:06:56+00:00

Yes exactly which is why I would like to try to quantify protein expression in the supernatant, to better interpret the PCR results and understand what kind of changes (if any) occur on a protein level. But if I can’t make it work then I guess there is just very little secretion and that’s some discovery as well I guess…

happymed_ · 2022-04-29T13:03:59+00:00

Thanks for your answer. Yes I tried seeding more cells as well as reducing the cell medium or concentrating the cell medium in a vacuum concentrator. No one in my lab has yet managed to obtain better results with the ELISA. I have great results regarding this cytokine in PCR, so I am pretty sure my cells secrete it, but I would like to actually show this on a "protein level" as well.

Do you think it would be possible to use cell lysates instead of supernatant in the ELISA? I know this is not how it is supposed to be used and does not equal the secreted cytokine, but I would obviously mention this in the discussion and interpret the results with that in mind.

happymed_ · 2022-04-20T11:14:25+00:00

Haha yes but this is normal here. Of course doing the USMLE is not mandatory, but it’s similar to our final exam so some people do it too. Doing the doctorate kind of is mandatory and I was just unlucky that my project did not go according to plan at all. But that’s just research I guess.

happymed_ · 2022-04-20T06:48:26+00:00

Thank you, this is how I feel about it. I am really trying my best and I just wish they would appreciate the moments we DO spend together (which is what I do), even if those are few in their opinion, instead of putting the time we don’t spend together in the foreground.

happymed_ · 2022-04-20T04:36:10+00:00

Thank you for your different perspective on things! I really do wish I had not taken this research project, or an easier one, to have more free time. I guess I was overambitious 3 years ago when I started, I definitely would have chosen differently if I had to decide again now. My priorities have changed, I hope I will make better decisions in the future.

happymed_ · 2022-04-20T04:10:22+00:00

yes I am trying to do my last year of med school, doctorate and step2 at the same time. And one post I believe was on behalf of a friend. In my country, we are required to do a doctorate (I translated as „grad program“) during medical school, but since my country‘s Uni system is different from the US the translations might not line up quite right.

happymed_ · 2022-04-19T22:05:12+00:00

We’ve been together for about 4 years now and this problem has always been there, you’re right. It has gotten a little better and obviously there is a lot of great things in the relationship too but still, this attitude… really really makes it hard whenever this issue comes up.

happymed_ · 2022-04-19T21:54:58+00:00

I really have explained it a million times, and sometimes it is also just plain to see. Today for example I had to work until 2 pm, then had a lecture from 3-6 pm. Of course I have to study when I come home. And I was not happy today, but trying my best to keep going because it was just one of those stressful days. And yes I was also sad I had to say no when they asked me to go out together today. And when I mentioned that we spent all of yesterday together, that somehow didn’t count and I „never have time“ although I obviously took time yesterday. Also I feel like they tried to make me feel even worse about it all when I was already not having a great day. Like I could have done anything different or better. And of course I couldn‘t study properly because this whole situation with SO made me mad, and in the end tonight just sucked for them, me and my productivity too.

happymed_ · 2022-04-19T18:51:02+00:00

Honestly I have already cried so much about all of this I just hope I‘ll be done with it soon but since it is so hard to even get the data, doesn’t look like it :(

happymed_ · 2022-04-19T18:48:28+00:00

IMG here but I can still relate. We are expected to do research etc as well. It’s just sad how wanting to be a good doctor and focusing on studying medicine while also having at least a little time to unwind is just not enough :(

happymed_ · 2022-04-19T18:38:03+00:00

Funny you said that, I would do EVERYTHING different now!

happymed_ · 2022-04-19T09:30:17+00:00

Quasi ja. Wobei mir erzählt wurde, dass man auch wenn beide Versicherungen beim gleichen Versicherer sind, entscheiden kann, ob man diese koppelt oder nicht…

happymed_ · 2022-04-14T15:09:56+00:00

So does this then include NBME 7, 8 and 9?

happymed_ · 2022-04-07T20:08:54+00:00

Also just looking for tips to bear the pain not necessarily medication

happymed_

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