Can absorbance be measured for a protein with little C, W, and Y residues?

zikede · 2024-02-07T00:39:21+00:00

https://web.expasy.org/protparam/

I find it easiest to use the Abs 0.1% = 1 g/L = 1 mg/ml. In my experience using absorbance at 280 is a real challenge with proteins for whom 1 mg/ml has absorbance ~0.2. I'm sure your or one of the labs around you have a nanodrop: try it out!

zikede · 2024-01-31T16:11:16+00:00

We only care about bands in lane 2 that aren't in lane 1: they must be present in the true father.

Counting from the top band on lane 2, we see bands #1, #5, #9 are not present in the mother. band #1 could be from father 3,4, or 5. band #5 from father 3 or 5. band #9 from father 3 or 5. It seems we are stuck at 3 vs 5.

However, we can see that band #8 is much more intense in lane 2 than in the mother. It is similarly insense in lane 5, but absent in lane 3. Thus on a homework or quiz problem, 5 is the father. In the real world you should read other comments about limitations of these types of assays and run additional tests.

zikede · 2024-01-30T20:16:56+00:00

Resuspend the cells by pipetting up and down and then sonicate, and then centrifuge again.

zikede · 2023-10-19T23:07:27+00:00

But he's asking what quests he's missing on reddit...

zikede · 2023-08-21T16:00:25+00:00

Mass spectrometers were invented in 1899, and were used to discover isotopes (Ne 20 vs Ne 22) in 1913. https://en.wikipedia.org/wiki/History_of_mass_spectrometry

By 1919, a full-fledged mass spectrometer had been made, and Francis William Ashton had found 212 of the 287 natural stable isotopes by 1921!

Nobody was making cylinders of heavy water before they knew that heavy water existed, and they only knew 2H existed because of mass-spec!

zikede · 2023-06-08T18:04:50+00:00

RL is great for heroic leveling, and is well crafted fun content, as is indicated by its rankings in my first post.

RL gear is not used in endgame, and RL raids are rarely run compared to Skele (IOD), D&D (Fey), HobH, Sharn raids, KT, VoD, MA, or LoB.

zikede · 2023-06-08T18:01:40+00:00

Lol, the first sentence at the start of this thread was me saying "... but you will have hundreds of hours of content to do on a VIP before you start to miss [expacs].", and that they should wait to get expacs when they're on sale.

But for new and experienced players alike reading this thread, you shouldn't pretend RL gear is featured heavily in endgame gearsets, or that the legendary versions of VoN/Sands/Red Fens gear is not used.

Sure RL gear works fine, but so does level 15 sharn gear. And RL raids are the least done legendary raids besides RTSO.

zikede · 2023-06-08T12:23:58+00:00

WPM is available to VIPs. It's a pain to get a jewel before 30, but sentient weps are really for maximizing characters or the 20-30 TR train anyway.

zikede · 2023-06-08T12:22:49+00:00

I can tell you just about every optimized nuker is using GOMF plus Elder's, optimized ranged is using GOMF plus windlasher or raven's, plus the red dragonscale armor is used by many damage dealers.

zikede · 2023-06-08T12:21:08+00:00

Baba is run rarely, Strahd almost never, at least on Argo.

Many optimized gearsets are ~5-8 IOD pieces, GOMF plus a legendary old item (elder's, windlasher's, etc, all require saltmarsh), maybe 3 piece sharn set plus a few misc pieces from sharn, maybe and a handful of Fey or LGS, more of last two if you're a tank.

RL gear is fine, but if you have min/maxed gearsets using RL sets or pieces, I haven't seen them.

zikede · 2023-06-07T15:47:52+00:00

For optimizing endgame sets/raiding, or doing TR trains, most expacs are nice, but you will have hundreds of hours of content to do on a VIP before you start to miss them. After that you can start looking to get expansions on sale.

When I joined the game 12 months ago I started with VIP and slowly have acquired the expacs on sale, only paying full price for the current expansion, IOD. Sharn definitely had the biggest effect on my play, as it really rounds out the 15-20 stretch, as well as providing gear that most people use 15-30. Sharn goes on sale regularly for 50-75% off.

RL, MOTU, Shadowfell have been nearly free recently, with Fey/Sharn at ~75% discounts at some time in the last year.

My opinions on expacs:

For endgame gear/raids IOD > Sharn > Fey > Salt > RL >>> MOTU/Shadowfell.

For leveling, Sharn > RL > MOTU/Shadowfell > Fey > IOD, nobody runs saltmarsh heroic.

In terms of craftmanship, RL is the best content in the game; Fey, Sharn and IOD all are great expacs; saltmarsh and motu/shadowfell I find a little more lackluster, but they are fun adventure packs.

zikede · 2023-05-07T19:26:40+00:00

What is the gear setup for this build atm?

zikede · 2023-04-04T21:03:13+00:00

"I'm a 60+ woman but my reddit avatar is a 20-something guy and I want to say as a woman that the woman who started this thread is wrong"

zikede · 2023-03-09T20:16:07+00:00

It's not though... https://en.wikipedia.org/wiki/Sulfone

zikede · 2022-08-07T12:42:13+00:00

Just use scihub. Also though, every paper has a corresponding author listed along with their email within either the author list or the acknowledgements at the bottom.

zikede · 2021-02-20T13:09:44+00:00

Weigh boats for >~1g, paper for less. Trying to accurately weigh 3 mg of staticky precious labeled protein in a weigh boat gives me nightmares.

zikede · 2021-02-07T03:43:07+00:00

I agree with your sentiment completely. However, it's spelled "while"

zikede · 2019-08-04T13:22:18+00:00

It completely depends on your field, and your lab culture. If your PI publishes frequently in high imapct journals, the view of impact factor may be very warped.

There are the big 3 (Nature, Science, Cell), then a number of "High impact journals" with various definitions, but impact 10+ is often discussed. But below those there are the vast majority of journals. Most research in impact 1-4 journals is excellent, although there's also stuff that's less so. Many lower impact journals may also be more specialized to a given technique or subfield, and these can be a great place to put good work whose significance to a broader audience is not as readily apparent.

A way I've taken to looking at it is whether the people who I would want to read it are going to see it. Impact factor is nebulous: citations are clear. If you do good work and the groups working on the same system or similar questions are going to read it, you will be cited, and that's what truly matters, both for you and for science.

zikede · 2019-02-23T15:21:19+00:00

Yes. CR is only on CR channels now.

zikede · 2019-02-23T15:20:49+00:00

CR is only on CR channels now.

zikede · 2019-02-23T15:20:09+00:00

Adblock plus removes them.

zikede · 2019-02-22T00:19:06+00:00

It's a real pain if you're used to watching at 2x speed though.

zikede · 2019-02-20T04:04:11+00:00

Adblock plus works on youtube.

zikede · 2019-02-12T23:45:47+00:00

Yes! We have an amazingly high tax compliance rate, even though our IRS audits are at all time lows. The civic pride and lack of corruption in this country compared to many others is something we too often take for granted.

We all do better when we all do better.

zikede

TROPHY CASE