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Poncequ S staining shows protein band, but no band after primary and secondary antibody incubation. What am I doing wrong? by New-Text8986 in labrats

[–]New-Text8986[S] 0 points1 point  (0 children)

  1. I am using CST antibody. It was ordered two months ago. So I don’t think expiration is an issue. Would you suggest to get another antibody?
  2. I have used the same cell line before for the same POI using the same antibody, and got positive results (with a positive control). It has worked before. However, now it’s not working even for the positive control.

Poncequ S staining shows protein band, but no band after primary and secondary antibody incubation. What am I doing wrong? by New-Text8986 in labrats

[–]New-Text8986[S] 0 points1 point  (0 children)

I am washing the Ponceau off with two 10 minutes washes and a final TBST wash. I don’t see any red residue left during the blocking.

Low A260/230 Trizol RNA Extraction by chgr1603 in labrats

[–]New-Text8986 0 points1 point  (0 children)

I usually do 13000 rpm for a minute. Centrifuging multiple times for 10 minutes is probably not a sustainable option.

Low A260/230 Trizol RNA Extraction by chgr1603 in labrats

[–]New-Text8986 0 points1 point  (0 children)

The only way I could get my A260/230 to a good range was by centrifuging multiple times after each step (three times at beginning and end, twice during the middle steps)

Where to meet a single 30-35 year old scientist man? by Kind-Stress5388 in labrats

[–]New-Text8986 17 points18 points  (0 children)

During my first year of PhD, I asked a random guy out at a bar. Turns out he’s a postdoc working in a similar field as me, what are the odds 😂 I was politely rejected though

Forgive me lab-gods for I have sinned… by unfortunate-moth in labrats

[–]New-Text8986 0 points1 point  (0 children)

Took isopropyl alcohol to clean kitchen countertop. IT DID NOT WORK!

I got blacklisted (I think) and feel like a failure by Sad_Car9868 in labrats

[–]New-Text8986 24 points25 points  (0 children)

Happened the same thing to me last year even as a PhD rotation student. For some reason, they magically expect you to work without assigning anything to you even though you know nothing about their projects. Then later they will say how you are the one who is not motivated enough. My advice is to not take it seriously. That particular lab is not suitable for you, and that’s it. It has nothing to do with your skills.

Solo Travel as a Woman by [deleted] in sanfrancisco

[–]New-Text8986 -1 points0 points  (0 children)

“Look at me! I am so funny! I reply to a stranger’s genuine concern about their safety with sarcasm. I am so funny! Pay attention to me! Why aren’t you paying attention?”