Purifying a new N-His6 tagged protein in the lab, and I've run a decent-ish Western blot with all of my fractions (soluble, insoluble, etc.) to check for solubility, concentration efficiency, etc. that comes with finding out where a protein ends up.
My Western (blocked with BSA, blotted with anti-His6, stained with CN/DAB) shows that my protein (red arrow) exists, but also I have some pretty noticeable bands at lower MWs. I am fairly confident that the three most prominent lower bands on the leftmost lane are proteolysis products that retain the His-tag. The faint higher MW bands I am also fairly confident are non-specific staining.
Are the fainter lower MW bands background bands due to overstaining and/or non-specific staining? How can I get rid of those in the future? I was thinking decreasing antibody concentration, better and more thorough washes before staining, and maybe a better blocking step? Any help would be appreciated!
Having the catalogue number under what category would this information normaly be found?
These things are kicking my ass and it takes a small stupid mistake to ruin a day and a half of work.
I've had battles off and on for my adult life with alcohol but I'm going through some intensive changes and stress lately. I've been drinking to deliberate blackout almost every night for 2 months now. I get what I have to get done during the day and then I just literally want to disappear. I'll pound multiple ginormous shots consecutively, black out, pass out and then get up to get done what I have to do. Repeat.
I don't know how to find an alternative for literally just wanting to disappear and not be conscious for a chunk of my life. I don't know what I'm really asking or what I expect from this post but I know I really need to stop doing this.
Hi all, after seeing the feedback on my previous adventure based on The Eagle's, Hotel California, I decided to write up another one.
This one is entirely based on The Rolling Stones': Paint it, Black.
Short Pitch: The town of Dartford is a charming enough town. The skies are always blue, the birds are always singing, and the death of the Lord's wife threatens to bring about the End of the World...? Lead your Party through a complex mystery, which culminates in a do-or-die battle to save the World from eternal darkness.
- Target Party: 3 to 4 Level 10+ Players
- Expected Playtime: 8+ Hours
- Tone: Mystery and Investigation; Desperate Fight for Survival
The Homebrewery link can be found below. Bear in mind that it is not pretty in the slightest. I am an amateur with the service, and this was my first real attempt at using it. A full synopsis can also be found below.
As always, leave comments, suggestions, and critiques down below.
Link: https://homebrewery.naturalcrit.com/share/BINnHFML5exT
A note on these adventures: These are not meant to be modules that you follow to the letter. I don't really intend for anyone to follow these exactly as written. Instead, my hope is that you all get something interesting from them to use in your own campaigns! Feel free to change names, encounters, stat blocks, entire plot points, characters etc. I really just like helping people brainstorm ideas :)
General Overview
Looking for work, or perhaps just a place to rest, a Party of Adventurers stumble into Dartford, a small hamlet. Quiet, colorful, and charming, the tiny village gleefully accepts the travelers. However, an overwhelming evil swells behind the scenes, threatening not only Dartford, but the entirety of the World itself.
- Target Party: 3 to 4 Level 10+ Players
- Expected Playtime: 8+ Hours
- Tone: Mystery and Investigation; Desperate Fight for Survival
This campaign draws its influence from the famous song, Paint it, Black by The Rolling Stones. If you haven't heard it (have you been living under a rock??) go ahead and give it a listen. The quest revolves around the town of Dartford, a small settlement under the direction of the Lord, Mikael Jaegar. This campaign begins slow enough. A murder here, a demonic beast there. However, clues found during these side-quests suggest a connection between these seemingly unrelated occurrences. Even more worrisome, the
... keep reading on reddit ➡I saw this short film in the early 2000s on the CW. It looked/felt like a student film project. A group of (4?) friends go exploring in a cave and find this stone bowl full of blood. They accidentally knock it over and the sun goes out. It starts getting really cold, and the group realizes they need to refill the bowl with blood again to get the sun back. They argue about who to kill until someone ends up accidentally dying, and I think they cry while they refill the bowl with his blood. The sun comes back and they look horrified.
Does any of that make sense? I haven't been able to find anything along these lines. Thanks in advance!
I’m roleplaying a character who fulfills the Tyranny of the Sun prophecy by permanently blotting out the sun. I’ve been looking around for SE mods that could fulfill this but many seem to be buggy or require you to use a power daily. This is a big problem since I am using Sacrosanct, and there is an endgame perk that makes sunlight instantly destroy you, so I can’t just recast the power daily (since I’ll just die immediately), it has to always be visually and mechanically active during the day. Any help?
Blot Out the Sky - XWB
Sorcery
>Create X tapped 2/1 white and black inkling creature tokens with flying. If X is 6 or more, destroy all noncreature, nonland permanents.
A sidegrade to [[Martial Coup]] this card is quite weird. Although most of the weirdness is from it's art. I'm sure it can find a use in some weird Orzhov tokens deck.
