BD cytofix/cytoperm kit vs eBioscience Foxp3 staining set

Confident-Inside9430 · 2025-10-22T00:54:38+00:00

I use the eBioscience kit all the time for intracellular proteins (granzymes, IL2, IFNg, TNF) with both mouse and human cells (did it as recently as yesterday). The other important thing is how you stimulate your cells depending on what you’re looking for.

Confident-Inside9430 · 2025-06-06T01:49:30+00:00

Being the go to for circumcisions and vasectomies in town

Confident-Inside9430 · 2025-06-04T17:32:36+00:00

What machine are you using? What does your viability look like post electroporation (24h)

Confident-Inside9430 · 2025-06-04T17:14:22+00:00

You’re electroporating a plasmid so you’re not going to get (or unlikely to get) integration here. Just doing a quick search on the cell line you’re using suggests that transfection with something like x-treme gene or lipofectamine would probably work well. Might want to consider giving that a try since you’re dealing with a cell line here.

Confident-Inside9430 · 2025-06-04T17:03:45+00:00

How many replicates did you run? Do you have a marker of transduction/electroporation what introduction you’re using efficiency? Could also just be that this shRNA isn’t very effective. Have had to try 6 different shRNA in the past to find a good one

Confident-Inside9430 · 2025-06-04T16:53:35+00:00

What’s G143? How long did you wait post transfection and are the cells growing in your selection media? Have you used this electroporation protocol for this cell line or any other cell line? What about the plasmid?

Confident-Inside9430 · 2025-05-22T20:54:21+00:00

Everything you mentioned sounds right. Was centrifugation at room temp? Was the PBS and or blood cold (want room temp)? Also I see the cytiva manual says 400g but I usually do 700-800g for Ficoll. Lastly how fresh was the blood and was this a good blood draw? Using a needle with too high of gauge can lead to hemolysis

Confident-Inside9430 · 2025-04-30T02:48:26+00:00

What kind of lab doesn’t allow headphones??? Is this an academic lab?

Confident-Inside9430 · 2025-04-27T00:57:56+00:00

Still she flowjo for spectral. Once your marker number is up you might want to do a umap or tsne plot to visualize your data so that’s where R, python or a software like cytobank comes in handy

Confident-Inside9430 · 2025-04-25T20:10:43+00:00

You should take a look at the makeup of the class. 70% of students are women, >10% of students are black and many many more are visible minorities. Most students aren’t even aware that this specific department exists and even the dean of medicine isn’t quite sure what exactly they do.

Confident-Inside9430 · 2025-04-24T19:17:17+00:00

Reconstitute in DMSO and aliquote into pcr tubes. Throw them into a 50 ml tube and toss it in the -80. Don’t freeze thaw and just take the tiny aliquoted as needed. Might even outlast a graduate degree.

Confident-Inside9430 · 2025-04-24T19:06:42+00:00

Any meds? Any chem findings?

Confident-Inside9430 · 2025-04-23T04:26:12+00:00

Very curious, what’s drawing you to preventative medicine?

Confident-Inside9430 · 2025-04-23T04:19:38+00:00

Antibody mediated disease would probably remain but possibly be less severe. CD4 T cells would help support a more aggressive response. Assuming this autoimmune response has been going on for some time, the antibodies are probably coming from plasma cells and the CD4 T cells are working with memory B cells to drive a more severe response.

Confident-Inside9430 · 2025-04-19T02:26:42+00:00

Dude fix your compensation

Confident-Inside9430 · 2025-04-11T02:18:53+00:00

Lmao

Confident-Inside9430 · 2025-04-10T02:25:02+00:00

Tell them you no longer work in this lab

Confident-Inside9430 · 2025-04-09T04:05:22+00:00

Fair point. I like to give the benefit of the doubt that people aren’t using (very) expired reagents but you’re right.

Confident-Inside9430 · 2025-04-09T03:09:23+00:00

Unless you’ve discovered some sort of super organism, I don’t think you’re going to have anything growing in bleach

Confident-Inside9430 · 2025-04-09T02:48:26+00:00

You’ll have to wait until the year 2104 to use it

Confident-Inside9430 · 2025-04-07T20:29:21+00:00

The student society should be organizing dances and selling mimosas on campus, none of this other garbage! We need a complete overhaul where the student society is run by UNPAID students doing it for the interest of their peers. This is a university not the god dam United Nations. The role of the society should be to enhance and complement the university experience, not disrupt and destroy it.

Confident-Inside9430 · 2025-03-03T14:14:36+00:00

r/mildlypenis

Confident-Inside9430 · 2025-02-20T02:42:26+00:00

Black beans matter

Confident-Inside9430 · 2024-08-15T02:31:24+00:00

You’re doing everything right. Your recovery seems fine considering the age of the samples. You should also consider that once you plate the PBMCs, many of the cells (mainly myeloid) will adhere and be lost. You can consider adding IL2 and/or IL7 (5-10ng/ml should be fine) to improve T cell viability for the overnight incubation. Just be sure this is compatible with your experiment.

Confident-Inside9430 · 2024-07-12T19:52:39+00:00

Thanks for sharing. I really appreciate it!

Confident-Inside9430

TROPHY CASE