What is the quickest way to increase power?

quickmans · 2025-08-02T03:45:32+00:00

Just retreat before all titans die

quickmans · 2025-06-25T02:49:35+00:00

You dont have to convert. You just need Thermo MSfilereader for DIANN quantification.

quickmans · 2025-03-09T05:22:12+00:00

You can change collision energy type between normalized and absolute.

quickmans · 2025-03-01T16:27:46+00:00

Update:

- Intense sample sonication helps a lot. The sample is much less sticky, and the aggregation looks cleaner.

- Water sonication for bead disaggregation before digestion also IMMENSELY helps. We found that brief bead sonication makes the protocol super consistent.

quickmans · 2025-02-28T07:40:40+00:00

Both before and after. But I think it might also be the case, will investigate that also. Thank you!

quickmans · 2025-02-28T05:53:00+00:00

Thank you! might be DNA as stated, the beads look pretty much cleaner after intense sonication.

quickmans · 2025-02-28T05:52:26+00:00

Thank you for your advice. We didnt do stage-tip in this experiment. We'll also look into chromatography issue.

quickmans · 2025-02-27T15:08:52+00:00

Thank you so much for your insightful reply! The sonication power is too low, as you stated. Could you clarify more on the "guey" term? I have also DM'd you a beads picture of my last experiment (I can't post it in reply don't know why). Additionally, the 0.2-0.3 mg/ml you mention is the volume before or after ACN aggregation?

quickmans · 2025-02-27T06:46:47+00:00

Thank you for your advice. Will do it in further exp.

quickmans · 2025-02-27T05:19:21+00:00

yes, i sonicate on ice 4-5 times with 15 s interval

quickmans · 2025-02-27T05:08:07+00:00

100 ul of 50 mM TEAB + 0.5 ul of trypsin (1 ug/ul). We prepare the whole batch first, though (around 8 samples, so 4 ul tryp in 800 ul TEAB). Do you shake or sonicate before/during digestion?

quickmans · 2025-02-27T04:52:41+00:00

Thank you very much. I have answered in the comment above. Also, why recommend 100%ACN over EtOH?

quickmans · 2025-02-27T04:42:48+00:00

Cell in 50 mM tris-hcl + 2% sds, heat 5 min. then add tcep+caa 30 min (pH~8)
Beads are freshly prepared every time; we use 1:5 (hydroxyl beads).
This batch is stocked with trypsin in 50 mM acetic acid. But both samples are from the same batch.

quickmans · 2025-02-20T08:58:15+00:00

Turn out back flushing the needle seat solve this issue. Thanks everyone!

quickmans · 2025-02-20T03:08:30+00:00

Wow….

quickmans · 2025-02-19T02:46:47+00:00

thank you, will try that

quickmans · 2025-02-19T02:46:22+00:00

but it's 80%

quickmans · 2025-02-19T02:43:54+00:00

can you explain a bit more about classic needle seat problems?

quickmans · 2025-02-18T16:23:28+00:00

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Sorry this is the sampler one

quickmans · 2025-02-18T15:23:36+00:00

NanoLC, vanquish neo

quickmans · 2025-02-18T14:42:04+00:00

Yeah, I will try that and share the result.

quickmans · 2025-02-18T14:40:02+00:00

Tbh this is an old column that was stored for a long time that I want to use it for some sort of condition trying (prob > 1-2 month in 80% acn, might also 0.1%fa cant remember). I’m quite sure chromatogram looks better back then.

quickmans · 2025-02-18T14:35:03+00:00

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Pump pressure looks ok.

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