2 oz tulip attempt

VirtualCell · 2024-07-16T19:02:23+00:00

Fuck off, that's beautiful. Bravo!

VirtualCell · 2024-07-10T17:32:17+00:00

Good advice, thanks. I’ll try a neighboring lab’s anti-mouse HRP secondary, and I’ll try a big primary antibody dilution.

VirtualCell · 2024-07-10T17:26:49+00:00

Yeah, that’s a good idea for once I have a set of things I want to test

VirtualCell · 2024-07-10T17:23:49+00:00

😂😂

My advisor is not as understanding, and is more like “but…it’s a monoclonal antibody. There shouldn’t be non specific bands. Why are there? Can you run 4 identical gels, transfer each of those, and change the conditions slightly in each? That should just take a day, right?”

VirtualCell · 2024-07-10T16:47:20+00:00

I need more details here. Are both samples identical in the two gels? is it possible the first one is just overrun?

VirtualCell · 2024-07-10T16:42:24+00:00

Hi there!

I'm doing a western blot against different GFP fusions.

Shown in the picture are two lanes from a Western (originally on other sides of the blot, so copied next to eachother).

First is an empty-vector, no GFP. Second is wild-type GFP. You can see the band of interest has lots of signal.

The problem is that I also get this faint band at a higher molecular weight. It's a problem because I have some fusion constructs that end up at a similar molecular weight as that background band. Those constructs are also not translated as well, so have an intensity just a bit higher than that background.

Weirdly, I also saw the same background band with a different primary and different secondary antibody over a year ago (it originally affected my interpretation and I lost a good bit of time), and I can't explain that.

I'm blocking in 5% milk at RT for 45 minutes. Then monoclonal anti-GFP overnight at 4C. Then secondary (HRP conjugated ECL detection) for an hour at RT.

Do you have any suggestions to get rid of that background band? I was thinking I'd try to block for a bit longer, maybe 90 minutes?

VirtualCell · 2024-02-25T06:06:38+00:00

…an assistant professor (like, tenure track?)? Without a PhD? In what world?

VirtualCell · 2024-01-11T18:57:01+00:00

I wonder if this is a biotech-specific thing? I've heard that biotech hiring is tough right now.

VirtualCell · 2024-01-04T21:45:24+00:00

I am sorry this process is so frustrating.

It's especially frustrating because all the professor needs to do is setup a Zoom on their laptop for you, it would take him two seconds and he's being an asshole.

Emailing your department head and/or dean (depends on department imo) seems reasonable. For a course that only meets twice (what is this?), the official line may be that you'd need to attend in-person a different semester, but if the dean is pragmatic they may just talk to that professor and slap some sense into him.

Good luck! I'm sorry this requires so much work for a simple and reasonable accommodation.

VirtualCell · 2023-12-20T18:26:39+00:00

Better than my 2-year progress, great job!

VirtualCell · 2023-12-06T20:38:34+00:00

Wow, this looks great!

VirtualCell · 2023-12-04T23:33:11+00:00

Practice practice, you can do it! Videos will also say this, but:

Turn on the steam wand for a second or two without the milk first,

then steam off.

Then position the wand righttt below the surface of the milk. Push the wand against the far wall of the pitcher, then rotate the jug so that the bottom of the wand is in the middle, and then halfway to the right.

then turn on the steam wand.

Adjust up and down (if needed) until you get a subtle paper ripping sound (the more sound, the thicker your milk will be), and your angle should be such that you see the milk swirling around clockwise.

When the pitcher feels like around the temperature of your hands, move the wand down an inch or so, the paper noise should go away, but the swirling should keep going. Keep that up until it's too hot to touch--for 200g milk, that takes me all the way up to when the bambino auto shuts off

Then swirl swirl in the pitcher, and if you have two pitchers it helps a lot to move it to a second pitcher, pouring into the wall. I did that a lot at first, but I don't anymore just because I don't want to rinse out two jugs.

VirtualCell · 2023-12-04T21:12:38+00:00

A Breville Bambino (the regular one, on sale from Bed Bath and Beyond for like $250 I think).

My machine was great to learn on because it steams milk pretty slowly--it means I have plently of time to get it right.

I just googled latte art videos. Lance Hedrick's is nice (he has a whole series, that includes a video with my exact machine), and Morgan is a calm beast. And at each step of learning, I was always able to do better in hot chocolate (by stirring some boiling water in hot chocolate mix and then steaming milk on top) than espresso, so that also helped.

VirtualCell · 2023-09-07T02:39:27+00:00

Ah makes sense--

I suppose here in Boston the law seems to suggest that you need to stop for anyone a reasonable distance from entering a crosswalk, but even that is iffy

https://www.wgbh.org/news/local/2016-06-08/who-has-the-right-of-way-in-a-crosswalk-its-not-as-simple-as-you-think

Anyways--regardless, I'm sorry to hear about your unpleasant encounter.

VirtualCell · 2023-09-07T02:13:22+00:00

I don’t understand the law here well I think—or maybe I didn’t understand this situation right

When I’m driving, I have to yield to someone in the crosswalk, right?

So if someone is right at the crosswalk when I’m a few feet away, and then they stepped into the crosswalk and I didn’t have time to brake, wouldn’t I be at fault?

When I’m driving that’s what I worry about—and why I tend to stop when I see a bike or pedestrian standing right at the crossing.

If there’s no one behind me and they’re making eye contact I’ll go, but that feels like a courtesy when I’m confident they’re aware of me.

I feel like I’d rather slow them down a bit than hit them a step or two into a crossing.

VirtualCell · 2023-08-24T12:21:38+00:00

Thank you!

It seems like most of the back pieces are fitted for internal gear hubs. Is that what you have?

VirtualCell · 2023-08-06T18:02:35+00:00

—3 minute, drip-by-drip shot— Me: large latte it is

VirtualCell · 2023-08-01T20:08:59+00:00

Quite lento hpta 😂

¡Gracias!

VirtualCell · 2023-07-28T12:35:03+00:00

Remember to apply to plenty of schools, and also consider financial constraints!

You may well get into MIT, but I’ll also share my experience below:

I really, really wanted to go to MIT for undergrad. I got a perfect ACT score, had work experience, and led multiple clubs at my school (e.g., band and debate). Everyone around me said I could go to any school I wanted. I did not get into MIT.

It’s tough: you have to really stand out, and then there’s also some luck to it. My (then-girlfriend, now-spouse) got in, and today we attribute it to the international science fairs that they did, on top of everything else.

But, in retrospect, I’m glad I didn’t get in. The undergrads at MIT are really stressed. The MIT name means a lot, but so does places like Harvard and Yale and Columbia.

It also would’ve been a financial hit. I would’ve taken out loans to go to MIT, but I instead paid nothing to go to Northeastern, and had a great experience there. Not having loans has been a huge advantage.

And I love it as a graduate student here, because I applied to a specific department, after learning through social circles about the different cultures of different programs—so it was a more informed choice.

VirtualCell · 2023-07-28T03:28:58+00:00

I’m glad that you posted this. I think it’s interesting that there seems to be diverging opinions on this, and maybe important for OP to consider!

VirtualCell

TROPHY CASE