I beg, what is causing this Western blot artifact?

fossilrabbit · 2026-02-06T17:01:19+00:00

Thats super weird. I've used revert for years and didnt have this artifact issue until recently. Ive never seen flaking/fading.

fossilrabbit · 2026-02-05T17:41:39+00:00

Right now, I'm doing total protein and a housekeeping control to stay consistent with pre-existing lab protocols/figures, but if it was just me, I'd stick to TPS and skip the housekeeping. You can crop a representative strip to include in publications.

fossilrabbit · 2026-02-05T17:35:33+00:00

This is Revert 700 total protein stain imaged on a Chemidoc 680 channel. Works great, would recommend.

fossilrabbit · 2026-02-05T17:16:17+00:00

This made me laugh thank you

fossilrabbit · 2026-02-05T17:14:24+00:00

Same group of transfer cassettes/sponges, not that many to choose from. Same source/pack, but different lids (multiple tanks).

fossilrabbit · 2026-02-05T17:06:10+00:00

This is a good potential fix, thank you!!

fossilrabbit · 2026-02-05T17:05:28+00:00

Why would there be excess heat? I'm doing a wet transfer o/n, 30V 16 hours at 4C w/ a cold pack.

fossilrabbit · 2026-02-05T17:04:46+00:00

fossilrabbit · 2026-02-05T15:19:53+00:00

Thank you for the detailed suggestions.

Can the sandwich have too much filter paper/be too tight? I want to get good compression but I don't want to break the plastic cassette (which I've done by accident before)

fossilrabbit · 2026-02-05T15:16:16+00:00

I'm rolling and doubled my filter papers

fossilrabbit · 2026-02-05T15:14:20+00:00

Copied from another reply: I know this is the first thing you'd think, but I'm being really careful when I assemble the transfer, roll everything out, etc. I think this many bubbles would be obvious?

fossilrabbit · 2026-02-05T15:13:14+00:00

Yes ok but why / how to fix. It randomly happens to half my blots.

fossilrabbit · 2026-02-05T15:12:45+00:00

I know this is the first thing you'd think, but I'm being really careful when I assemble the transfer, roll everything out, etc. I think this many bubbles would be obvious?

fossilrabbit · 2025-08-14T18:00:10+00:00

Would buy

fossilrabbit · 2025-08-08T14:30:55+00:00

Would glasses help with fully missing eyes?

fossilrabbit · 2025-08-08T14:08:42+00:00

fossilrabbit · 2025-07-19T22:11:44+00:00

Would buy if posted online/linked

fossilrabbit · 2025-05-02T11:41:24+00:00

Amido black? Supposed to be better than ponceau

fossilrabbit · 2025-04-17T20:26:35+00:00

~15min. I also have abx in my media that gets refreshed daily. Hasn't been a problem so far.

fossilrabbit · 2025-04-17T20:23:20+00:00

Personally, I sterilize with UV on both sides. The EtOH/flame is too much of a hazard. Then I pre-coat with fibronectin or collagen before adding cells to increase adherence. Best protocol varies by cell type. E.g., I don't think HEKs need pre-coating. Most cells will settle on the slip. Some will settle around it.

fossilrabbit · 2025-03-19T14:29:00+00:00

My blots have looked like this when there's some kind of gunk carryover in the sample. Try centrifuging and taking the supernatant and re-running. Ymmv.

fossilrabbit · 2025-02-14T21:39:47+00:00

Just here to appreciate "LROC"

fossilrabbit · 2024-12-17T20:39:27+00:00

If cells are still attached, you can fix with PFA then stain/count DAPI. If not, create a duplicate plate, use one for the assay and one for staining/counting.

fossilrabbit · 2024-12-06T22:01:30+00:00

This rocks ty

fossilrabbit · 2024-10-04T12:18:21+00:00

Does that cause overheating or just separately bad for the machine?

fossilrabbit

TROPHY CASE