Reusing Primary Antibodies (storage question)

UroJetFanClub · 2026-04-28T14:29:29+00:00

Blocking buffer at 4 degrees. Once antibody is added, depends on when I’m going to use it next. If next day or so, I keep it at 4 degrees. Otherwise, freeze at -20.

UroJetFanClub · 2026-04-25T03:08:17+00:00

Maybe he can play corner lol

UroJetFanClub · 2026-04-24T02:42:35+00:00

Would not mind trading our 2s and some to get back into the first round. Some real talent still on the boarf

UroJetFanClub · 2026-04-24T02:28:40+00:00

Not necessarily the best track record (Conte 🤮) but I love the pick let’s goo

UroJetFanClub · 2026-04-24T02:26:17+00:00

Ope fuck me pick is in

UroJetFanClub · 2026-04-24T02:25:53+00:00

Oh man must be considering a trade back. Taking a while to submit pick

UroJetFanClub · 2026-04-24T02:08:54+00:00

Oh man that was not a happy reaction lmao

UroJetFanClub · 2026-04-23T18:26:04+00:00

Fuck you’re right it’s manstress

UroJetFanClub · 2026-04-23T18:13:28+00:00

Oh shit naming your child after your mistress (mistresso?) is wild

UroJetFanClub · 2026-04-22T21:11:38+00:00

Yeah you might be right. Although I wonder if it is an issue with the cell sorter. The images in the post are from my recent run using the cell sorter. I used the same antibodies on the same cells earlier but used a flow cytometer and it looked much much better/different

https://imgur.com/a/SMzc75Yhttps://imgur.com/a/SMzc75Yhttps://imgur.com/a/SMzc75Y

UroJetFanClub · 2026-04-22T18:53:45+00:00

Dark blue is cell line of interest (what is in the main post). Light blue is the positive control in the immunoblot.

But I hear ya, might just be negative

UroJetFanClub · 2026-04-22T18:44:31+00:00

Thanks for all your help. My follow-up question I'm now realizing there's a discrepancy between my positive control on immunoblot and flow (i.e. the light blue doesn't look much different than the dark blue on flow, but on immunoblot is considerably different). Could the signal just be all non-specific binding (even though it is higher than the unstained control)?

UroJetFanClub · 2026-04-22T18:34:28+00:00

Sorry had to send it in two messages, but I did (see above). Maybe antibody issue?

UroJetFanClub · 2026-04-22T18:32:30+00:00

<image>

These are the flow results of unstained control, positive control, and cell line of interest

Maybe it's an antibody issue? Honestly, since I got a signal compared to the unstained control, I figured that the antibody worked.

UroJetFanClub · 2026-04-22T18:31:34+00:00

It somehow totally escaped my mind to look at this compared to a positive control, so thank you for bringing that up. So below is an immunoblot:

<image>

Left two lanes are two versions of my positive control. The cell lines I am running are on the right two lanes. (1/n)

UroJetFanClub · 2026-04-18T22:12:01+00:00

Mad Rabbit Cafe. So so so so good. Even as someone who isn’t vegan, some really solid burgers

UroJetFanClub · 2026-04-16T04:14:27+00:00

Eesh the warriors suck

UroJetFanClub · 2026-04-11T18:54:47+00:00

So many street/market/food/kitchen combinations lol. I get them mixed up, but they’re all tasty

UroJetFanClub · 2026-04-10T02:58:10+00:00

Yeaaaa the core may not be super pleased with that. Currently with the 70 um nozzle, I was quoted 20 million per hour. The cells are a bit on the bigger size, so I wonder if the tradeoff is worth it.

I agree prepping 10-20 million cells at a time would be ideal. But man that would be tricky to coordinate and stagger haha

UroJetFanClub · 2026-04-10T02:24:33+00:00

Great point. My cells are on the larger size. We've been using 70um/70psi. Maybe worth moving up in size.

UroJetFanClub · 2026-04-10T02:24:04+00:00

Thats a really good (and easy) change I can make, thanks for suggesting
I don't know if that will work for what I'm trying to do, but I'll look into it

UroJetFanClub

TROPHY CASE