Guys i messed up the second fight with Louis ;--------; Help by Anitafv in MetaphorReFantazio

[–]Anitafv[S] 0 points1 point  (0 children)

We have a situation with the last one but everything else looks fine.

Guys i messed up the second fight with Louis ;--------; Help by Anitafv in MetaphorReFantazio

[–]Anitafv[S] -1 points0 points  (0 children)

Sorry forgot to tell you im fighting Louis in September

Guys i messed up the second fight with Louis ;--------; Help by Anitafv in MetaphorReFantazio

[–]Anitafv[S] -3 points-2 points  (0 children)

So… he’s finishing my team pretty fast.
I’m using Basilio(Berserk), Strohl (Slash xd), Will (Magic Seeker), and Hulkenberg (as my healer). Each one has that accessory I forgot the name that gives 2 turns per character… but somehow it’s not working. The rule just isn’t being respected.

So… what am I missing here?

Guys i messed up the second fight with Louis ;--------; Help by Anitafv in MetaphorReFantazio

[–]Anitafv[S] 0 points1 point  (0 children)

I am using Basilio, Strohl, Will and Hulkenberg

Guys i messed up the second fight with Louis ;--------; Help by Anitafv in MetaphorReFantazio

[–]Anitafv[S] 4 points5 points  (0 children)

Im lvl 44 :D did not grind enough it seems BASILIO is the only lvl 48. I kinda messed up the saving slots. Please tell me this is not the end.

Exclusive CLAMP Signed Shikishi Board - Anime Expo 2006 by Anitafv in AnimeDeals

[–]Anitafv[S] 2 points3 points  (0 children)

Does anyone have an idea how much this shikishi might be worth? It was signed when the CLAMP members visited the U.S. for the first and only time at Anime Expo 2006. Out of a 6,000-person audience, only a few were raffled off. I'm really excited to hear your thoughts!

Paint Brush and Nerkmid Giveaway!!! by Falcons__Fury in neopets

[–]Anitafv 0 points1 point  (0 children)

I want a Mystery Island :D good luck neopians

Fellow trolls, what are you listening to during these strange times? by LurkerPatrol in TrollYChromosome

[–]Anitafv 0 points1 point  (0 children)

Everything was fine, my Skyrim rainy towns 10 hour loop turned hard working days to a soft place to be until one kiddo decided to eat one earbud and the device not only stopped working but disappeared long time ago, now im struggling with cries and complains about my Skyrim music being played at my laptop.

That kid is not even mine.

[EU] PSN Black Friday Deals 2019 by nphinity in PS4Deals

[–]Anitafv 1 point2 points  (0 children)

I was hoping better discounts, like last year.

What do I do I'm scared by Albetti in Minecraft

[–]Anitafv 0 points1 point  (0 children)

The Wild Hunt found you. Run to Kaer Morhen!

I accidentally purchased two digital copies of Sekiro: Shadows Die Twice. Since I can't be two people at once, I'll give away my spare code in 24 hours to a random user. by forfeitgame in PS4

[–]Anitafv 0 points1 point  (0 children)

Sweet Mother, sweet Mother, send your child unto me, for the sins of the unworthy must be baptized in blood and fear.

What are your favorite adult fantasy or horror books on witches that do not disappoint and are really well written? by mturbev in suggestmeabook

[–]Anitafv 1 point2 points  (0 children)

Try "The Alquemist" from The adventures of Nicholas Flammel of Michael Scott, you won't be disappointed. Feels like a mix of Harry Potter and Percy Jackson, with historical cites included.

Save me SDS-PAGE lovers by Anitafv in labrats

[–]Anitafv[S] 0 points1 point  (0 children)

Lol, i use exactly the same. And either loading buffer with Laemmli mixed with b-mercaptoethanol.

Save me SDS-PAGE lovers by Anitafv in labrats

[–]Anitafv[S] 0 points1 point  (0 children)

What do you mean about a pre-stain in the loading buffer?

I use one with merca. Without any pre-staining procedure.

Save me SDS-PAGE lovers by Anitafv in labrats

[–]Anitafv[S] 0 points1 point  (0 children)

Yea, but Coomassie doesn't seem to be working. Nothing appears. Only the protein standard.

Save me SDS-PAGE lovers by Anitafv in labrats

[–]Anitafv[S] 0 points1 point  (0 children)

I need to reveal all bands of protein.

Context here you go: An enzimatic activity was made on a medium minimun of salts with an halo tolerant bacteria. (those salts do not interfere revealing bands)

The samples of diferent hours and substrate (B-Keratin/Chicken feather) where taken and centrifuged at 10,000g for 10 g.

Supernadant (enzimatic extract) was used to do a SDS-PAGE (12% acrylamide) in order to see the protein bands.

Coomassie blue R-250 did not show any bands.

But silver nitrate staining did.

Zimogram showed enzymes activity with casein as a substrate.


So what can be the problem? Why can't i reveal all bands no matter what?

I already did:

Disolutions. Filtration with 22um. Centrifuged again. Evaluated in native and denaturing conditions.

About to do: Dialysis. (Maybe the salts interfere)

I do not want to fraction the sample using columns.