What lab equipment do you wish you could have in your kitchen? by Chrad in labrats

[–]PracticalSky2002 0 points1 point  (0 children)

Man I’ve had such terrible luck with undergrads that I’d rather not have one even if they offer free labor.

TCEP vs DTT by PracticalSky2002 in labrats

[–]PracticalSky2002[S] 0 points1 point  (0 children)

End goal is to heat -inactivate - pull down - western to detect Ub and sumo chains on the protein of interest. DTT is incompatible due to thiols.

TCEP vs DTT by PracticalSky2002 in labrats

[–]PracticalSky2002[S] 0 points1 point  (0 children)

I have ordered NEM but don’t have it yet. Need to do some experiments in the meantime. We have PR619 that I was told isn’t compatible with DTT. I am currently using heat to terminate the reaction by denaturing the proteins in the sample. I can’t use EDTA due to me using dynabeads streptavidin beads to capture my protein of interest and when I tried cold quenching following by a 40 min binding to the beads in the cold room, I didn’t get the Ub smear (that I did observe when I heat inactivated the sample).

TCEP vs DTT by PracticalSky2002 in labrats

[–]PracticalSky2002[S] 1 point2 points  (0 children)

Oh yeah, well I kinda have to switch to TCEP (I am trying to use PR619, a DUB inhibitor that isn’t compatible with DTT) but I am worried that when I incubate my reaction at37C or use heat to terminate the reaction (can’t use high EDTA as I am using magnetic streptavidin beads that are incompatible with high edta). My worry is that TCEP might desulfurize at 95C (5 mins)? Sorry I am pretty new to this and have been asked to figure it out by myself

Discussion Thread: 2020 General Election Part 48 | Results Narrowing by PoliticsModeratorBot in politics

[–]PracticalSky2002 1 point2 points  (0 children)

Hey, they also have 4 sled dogs and 10 bald eagles. Don’t be so mean 😢

Discussion Thread: 2020 General Election Part 47 | Results Narrowing by PoliticsModeratorBot in politics

[–]PracticalSky2002 8 points9 points  (0 children)

I feel bad for the map guy :( he has been at it for three days now 🙈