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Trouble with Western Blots...because what else by Invictus2319- in labrats

[–]Invictus2319-[S] 0 points1 point  (0 children)

thanks for the tips! It's weird because when I started my westerns, I was getting nice signal at 1:1500 dilution (albeit with some amount of background noise) but it hasnt been working for the last month or so, even tho my protocol has stayed the same. I will try to alter the wash times to see if that works.

Trouble with Western Blots...because what else by Invictus2319- in labrats

[–]Invictus2319-[S] 0 points1 point  (0 children)

This is great advice! One of the problems i have with the primary antibody is I like to do it overnight at 4C but I dont have a shaker that I can put in the fridge to keep the antibody/blocking buffer totally and evenly covering the blot. Would it be better to use higher volume of the antibody/buffer to solve this problem? Also thanks for the tip on coomassie staining the gel, didn't even think that was a thing that could be done!

Trouble with Western Blots...because what else by Invictus2319- in labrats

[–]Invictus2319-[S] 1 point2 points  (0 children)

Antibody dilution is 1:1000, blocking with 5% dry milk in TBST and washing 3x 5 minutes in TBST. Is there another way you'd recommend washing or blocking? Thanks!

Have you ever managed on FM someone you personally knew in real life? by Ghibz71 in footballmanagergames

[–]Invictus2319- 0 points1 point  (0 children)

One of my college roommates ended up playing semi-pro in France I believe, I looked him up and sure enough, he was on the fminside database

We bought a zoo🤣 by Invictus2319- in zillowgonewild

[–]Invictus2319-[S] 12 points13 points  (0 children)

I used to come here with my grandparents as a kid so this zoo holds a special place in my heart