Protocol Online logo
Top : New Forum Archives (2009-): : SDS-PAGE and Western Blotting

Undergrad "activated" a NS membrane with 100% MeOH... Any hope? - (Jun/17/2013 )

Hey all,
So the undergrad tried to activate a nitrocellulose membrane with 100% MeOH. Is there any hope of getting an actual result with this now?

Thanks in advance for your help!

-wilson.2109-

Only time I did that by mistake the membrane disintegrated in the methanol so I'll say no. But I guess if the membrane has kept it's physical integrity you might want to try. Was this after transfer? If so, I guess you could ponceau stain to see if the proteins are ok and then go ahead or not. If this was before transfer, I'll say discard a cut a new piece.

just my thoughts :)

-almost a doctor-

You shouldn't be so hard on the undergraduate mistakingly activating the nitrocellulose in methanol. 1) You are coming to this forum to ask a very obvious question, 2) You obviously are not being clear enough with the student worker, 3) I'm sure you were not peaches when/if you were an undergraduate researcher. The methanol will have no effect on the nitrocellulose. Think about why methanol is present in your transfer buffer. Quit being a dick.

Edit:
easy google search

http://www.protocol-online.org/biology-forums-2/posts/11391.html

-jerryshelly1-

jerryshelly1 on Mon Jun 17 16:26:01 2013 said:


You shouldn't be so hard on the undergraduate mistakingly activating the nitrocellulose in methanol. 1) You are coming to this forum to ask a very obvious question, 2) You obviously are not being clear enough with the student worker, 3) I'm sure you were not peaches when/if you were an undergraduate researcher. The methanol will have no effect on the nitrocellulose. Think about why methanol is present in your transfer buffer. Quit being a dick.

Edit:
easy google search

http://www.protocol-...osts/11391.html


Thank you for your kind response jerryshelly1. I really appreciate you taking the time out of your day to help another person out. The membrane was partially disintegrated and I was not sure if there was any trick for saving it. So clearly the methanol did have an effect on the nitrocellulose membrane. And for the record, the reason I asked is because I do actually care about this undergrad and I was trying to save him from getting yelled at by our PI, because said undergrad was given specific instructions to not activate the membrane.
Quit spending your time being an internet troll, it helps no one, especially you.

-wilson.2109-

wilson.2109 on Mon Jun 17 23:37:44 2013 said:


jerryshelly1 on Mon Jun 17 16:26:01 2013 said:


You shouldn't be so hard on the undergraduate mistakingly activating the nitrocellulose in methanol. 1) You are coming to this forum to ask a very obvious question, 2) You obviously are not being clear enough with the student worker, 3) I'm sure you were not peaches when/if you were an undergraduate researcher. The methanol will have no effect on the nitrocellulose. Think about why methanol is present in your transfer buffer. Quit being a dick.

Edit:
easy google search

http://www.protocol-...osts/11391.html


Thank you for your kind response jerryshelly1. I really appreciate you taking the time out of your day to help another person out. The membrane was partially disintegrated and I was not sure if there was any trick for saving it. So clearly the methanol did have an effect on the nitrocellulose membrane. And for the record, the reason I asked is because I do actually care about this undergrad and I was trying to save him from getting yelled at by our PI, because said undergrad was given specific instructions to not activate the membrane.
Quit spending your time being an internet troll, it helps no one, especially you.


If the membrane was partially degraded would you not have the sense to throw it out. Nitrocellulose is a cheap consumable. Your initial tone was derogatory against the undergraduate. I am assuming you posted on this forum, immediately threw out the membrane and began again. I am sorry if I sounded like an "internet troll". I just dislike it when individuals use this forum for ranting, it helps no one, especially you.

-jerryshelly1-

One thing I have noticed over my years being on various forums is that what anyone writes is interpretable in many different ways.

For instance if I wrote in reply to something: "Yeah right, that would work." I could be being sarcastic, joking, or actually just answering in a casual manner, but you would never know which without further input from me.

Just something to bear in mind for both of you...

-bob1-