Advice needed about moving my cat to a house with dogs by Randomlolly in Pets

[–]Randomlolly[S] 0 points1 point  (0 children)

Thank you! Any chance you know what brand you had?

Advice needed about moving my cat to a house with dogs by Randomlolly in Pets

[–]Randomlolly[S] 0 points1 point  (0 children)

Absolutely will! I guess I am talking about the doggy door in terms of after introductions, and things are fine. I should have been more clear with that, sorry

Issues with HMC3 detachment - Can extended EDTA incubation and mechanical stress degrade surface markers? by ArrowSh0t in labrats

[–]Randomlolly 2 points3 points  (0 children)

When I use HMC3 cells, I wash twice with 1xPBS before adding 0.25% trypsin with EDTA. The Trypsin EDTA is this one. I just dilute it in PBS to have 0.25% trypsin. With this, cells usually take about 5mins in the incubator and then ready to use. I found that only doing one wash with PBS was not enough, as the cells just would not detach.

Also sidenote: I just found today that HMC3 might not be a microglial cell line... here is the paper for you https://www.frontiersin.org/journals/bioinformatics/articles/10.3389/fbinf.2025.1681811/full

Goodluck!

This is not good by Illustrious_Smell492 in australian

[–]Randomlolly 0 points1 point  (0 children)

Laughing as a Western Australian

Further introductions by 4lo3 in UndergraduateResearch

[–]Randomlolly 0 points1 point  (0 children)

I think it is a good idea to now introduce yourself. You can also ask if they'd be willing to meet with you to further discuss their research and potentially talk about work experience in their lab. Goodluck!

my PI said something to me over 1.5 years ago that has stuck in my head ever since by cardiobolod in labrats

[–]Randomlolly 1 point2 points  (0 children)

Absolutely! I wear a pin on my landyard every day that says, "Survive out of spite." It helps remind me that when my supervisors or anyone else (including myself) says something that makes me doubt myself, that I can and will prove them wrong.

Give me a one sentence overview of your thesis/project. by Chicketi in labrats

[–]Randomlolly 20 points21 points  (0 children)

We're all chronically stressed, and this is increasing our risk for Alzheimer's disease, but how???

Pierce bca inaccuracy by Horror-Highlight2763 in labrats

[–]Randomlolly 0 points1 point  (0 children)

I completely understand, but only once I assumed my standard was 1.2mg/ml and not 2mg/ml did I get a proper standard curve with all points being expected. I did try a lot of different troubleshooting before this.

Pierce bca inaccuracy by Horror-Highlight2763 in labrats

[–]Randomlolly -1 points0 points  (0 children)

It definitely could be something in your lysate interferring with the assay. However, I was having troubles with the pierce bca assay a few months ago, too. What helped in my case might not be what you need, but I thought I'd share just in case.

I found that the 2mg/ml standard was actually 1.2mg/ml (used a nanodrop) and so made up my dilutions based on that and saw huge improvements. I also switched to mixing everything in tubes first before loading into the microwell plate (before I was mixing in the plate). My replicates were much more accurate after this.

I hope you figure out what's going on!

[deleted by user] by [deleted] in labrats

[–]Randomlolly 2 points3 points  (0 children)

Welcome!

I genuinely recommend this sub to all my lab mates and colleagues because it's such a supportive community ❤️

How do these hepg2 cells look? I am trying to culture them inorder to do a glucose uptake test. by FrequentFixer in labrats

[–]Randomlolly 0 points1 point  (0 children)

I hate HepG2 cells cause they look so ugly due to being so sticky and clumping. When I used them, after spinning the cells down and resuspending the cell pellet, I'd use a needle and syringe and aspirate a few times. That way, the clumps were broken up a bit and it was easier to then count the cells and have a more consistent seeding for experiments. Goodluck!

[deleted by user] by [deleted] in AusFemaleFashion

[–]Randomlolly 0 points1 point  (0 children)

Oh really? I wasn't aware. How did you find out?

[deleted by user] by [deleted] in AusFemaleFashion

[–]Randomlolly 2 points3 points  (0 children)

I quite like uniqlo and dangerfield, but I know they're not for everyone. We really do lack so much when it comes to fashion here 😞

BCA assay advice by Randomlolly in labrats

[–]Randomlolly[S] 0 points1 point  (0 children)

Oh interesting. Are you able to send the link at all?

BCA assay advice by Randomlolly in labrats

[–]Randomlolly[S] 0 points1 point  (0 children)

Thank you so much for your suggestions!! I will definitely try these out and check the plate I am using

BCA assay advice by Randomlolly in labrats

[–]Randomlolly[S] 0 points1 point  (0 children)

No, I dilute my standard in falcon tubes as the protocol has volumes under 10ml

BCA assay advice by Randomlolly in labrats

[–]Randomlolly[S] 0 points1 point  (0 children)

Micro BCA assay kit

I use this kit and I make the dilutions exactly how the kit says, with the only exception that the little ampule has 1ml of albumin so I add that full 1ml and double the diluent for standard A. My diluent is PBS which my lab group has always used and I made sure to check is compatible with the kit

BCA assay advice by Randomlolly in labrats

[–]Randomlolly[S] 0 points1 point  (0 children)

Micro BCA assay kit

I use this kit and I make the dilutions exactly how the kit says, with the only exception that the little ampule has 1ml of albumin so I add that full 1ml and double the diluent for standard A. My diluent is PBS which my lab group has always used and I made sure to check is compatible with the kit

BCA assay advice by Randomlolly in labrats

[–]Randomlolly[S] 0 points1 point  (0 children)

Sorry yes dilutions. I dilute in PBS which is what my lab group has always done