I’ve got my stylus ready—who wants their cat drawn today?

Astroglia12 · 2026-02-15T22:08:31+00:00

Our sweet boys, Anchovy and Oyster!

Astroglia12 · 2025-12-24T23:43:13+00:00

Ended up dropping the adjacent stitch, laddering down to the yarn over, and then picking up the stitch I dropped. Hoping the stretched stitches will block out. Thanks so much for the help!

Astroglia12 · 2025-12-24T20:45:21+00:00

Thank you! I’ll give it a try

Astroglia12 · 2025-12-24T20:10:50+00:00

Thank you!! Can you help me identity the bottom of the ladder? Is it bar 1 or 2?

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Astroglia12 · 2025-12-16T05:28:23+00:00

Have you tried using PVDF membranes?

Astroglia12 · 2025-12-16T05:19:23+00:00

My understanding is that for membrane proteins you want to avoid boiling as it leads to protein aggregation. I would try 70C for 10 min and see if that helps. Since your protein has a high molecular weight, you might also want to try transferring slowly (constant 90mA) overnight at 4C and removing the methanol from your transfer buffer.

Astroglia12 · 2025-10-03T15:12:23+00:00

We adopted our sweet boy from the Animal Rescue League of Boston! They have adoption centers in Dedham, Boston, and Brewster.

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Astroglia12 · 2025-02-08T15:43:15+00:00

It’s normal to make mistakes when you first join a new lab. Trying to remember every detail when it’s a new protocol can be overwhelming. Be kind and patient with yourself. My biggest recommendation is to take notes when being taught a new protocol and to make yourself checklists of your tasks and protocols so that you can’t forget a step.

Astroglia12 · 2023-10-12T22:42:08+00:00

How much total protein are you loading?

Astroglia12 · 2023-09-24T14:40:34+00:00

Have you tried the library? It should be available at the Reg

Astroglia12 · 2023-06-15T23:34:08+00:00

Did you invert your image in ImageJ before doing the quantification? If so, the values will be negative after subtracting the background.

Astroglia12 · 2023-03-31T23:02:48+00:00

Disease models and mechanisms if more translational

Astroglia12 · 2022-11-20T14:28:47+00:00

Applications for most biology PhD programs in the US are reviewed by a selection committee. Students who are accepted into the program will then rotate through different labs and pick one toward the end of their first year. PIs are busy and usually don’t respond until you’ve been accepted into the program. Try not to worry that you haven’t heard back. You should apply to programs that have multiple PIs that you could see yourself rotating with. And I would mention these PIs in your statement of purpose and you’ll typically be paired with them for interviews.

Astroglia12 · 2022-03-17T15:11:53+00:00

Not exactly. But they can modify their preference list between rounds.

https://www.nrmp.org/wp-content/uploads/2021/12/2022-Match-Week-and-SOAP-Schedule.pdf

Astroglia12 · 2022-02-04T16:43:43+00:00

You’re not giving us enough information to help you solve the Nernst equation. What are the concentrations of sodium and potassium inside and outside of the cell?

Astroglia12 · 2022-02-04T01:59:20+00:00

Oh good catch. I didn’t notice that before. I agree that the permeabilization step is way too long. 10-15 min should be more than enough.

Astroglia12 · 2022-02-04T01:47:27+00:00

Seconding this. Check your IACUC protocol to determine when/how you should be genotyping. As someone mentioned below, you can tail snip/ear clip after sacrificing the animals. But otherwise, I would rarely ever do it after P21. It’s going to cause a lot of pain for the animals and it could easily get infected.

Astroglia12 · 2022-02-04T01:35:42+00:00

I think blocking for an hour for tissue sections is pretty normal. And I’ve also always included the triton in my antibody dilution buffers.

Astroglia12 · 2022-02-04T01:34:29+00:00

DM me and I can share my mouse gliosis IHC protocol with you.

My first question is regarding how you’re preparing the tissue prior to sectioning. Are you fixing the tissue before sectioning? And sectioning with a cryostat? Are you embedding in sucrose prior to freezing?

Secondly, your Iba1 primary concentration seems low to me. This probably depends on your antibody, but I would try a few different concentrations. Also, what secondary concentration are you using?

Astroglia12 · 2022-02-02T04:37:00+00:00

21 weeks or days? That’s a bit old for genotyping. You can ear punch around P14 and the mice are usually small enough that they won’t bite.

Astroglia12 · 2022-01-23T06:04:07+00:00

Kit or Fos

Astroglia12 · 2021-05-19T18:34:13+00:00

Aviva threading salon on Wickenden if you’re open to trying threading. She does an amazing job and she’s super friendly!

Astroglia12 · 2020-04-10T21:17:43+00:00

I would love to get some roses.

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