November 28, 2024, 05:48:50 PM
Forum Rules: Read This Before Posting


« previous next »
Pages: [1]   Go Down

Topic: Diagonal Electrophoresis  (Read 8618 times)

0 Members and 1 Guest are viewing this topic.

Offline Albert

  • Lonely Wanderer
  • Retired Staff
  • Sr. Member
  • *
  • Posts: 1050
  • Mole Snacks: +112/-22
  • Gender: Male
  • Half Decent Pharmaceutical Chemist
Diagonal Electrophoresis
« on: December 31, 2005, 08:20:02 AM »
Hello, everybody.
I hope everyone to have a nice New Year's Eve tonight!

However, before night comes, would you be so kind as to answer me?

I know detection of disulphide linkages (when we determine the primary structure of a protein) can be carried out with the help of mass spectrometry (MALDI spectra).
I read of a different approach based on a so-called diagonal cellulose acetate electroforesis. I just know you oxidize, with performic acid, SH-groups (only those which formed disulphide lingages) to SO4--, and then you perform your electroforesis.

The questions are:
- what does 'diagonal' mean in this situation?
- how do SO4-- groups 'behave' or how can you recognize them?

Thank you and have nice parties! :partytime2:
« Last Edit: December 31, 2005, 08:50:54 AM by Albert »
Logged

Offline Alberto_Kravina

  • Assault Chemist
  • Retired Staff
  • Full Member
  • *
  • Posts: 608
  • Mole Snacks: +70/-15
Re:Diagonal Electrophoresis
« Reply #1 on: January 02, 2006, 09:00:12 AM »
Hey Albert!

When you perform diagonal electroforesis you "split up" your sample in two dimensions.
The sample is splitted up in one direction, after that you oxidize with performic acid, and then you split up your samples vertically to the first direction of electrophoresis.

What I attached:

a) if your protein doesn't have any -SH groups you'll get a line of protein spots.

b.)If there are -SH groups in your sample you oxidize them to R--SO3- groups, which have a different behaviour. This proteins create some spots that are NOT on a line (I marked those spots with an arrow)

Hope this helps 8)
« Last Edit: January 04, 2006, 10:14:03 AM by Alberto_Kravina »
Logged

Offline Albert

  • Lonely Wanderer
  • Retired Staff
  • Sr. Member
  • *
  • Posts: 1050
  • Mole Snacks: +112/-22
  • Gender: Male
  • Half Decent Pharmaceutical Chemist
Re:Diagonal Electrophoresis
« Reply #2 on: January 02, 2006, 01:05:44 PM »
Quote from: Alberto_Kravina on January 02, 2006, 09:00:12 AM
Hope this helps

Not only does it help, but it also completely solves my problem.

Thank you very much.
Logged

Offline Alberto_Kravina

  • Assault Chemist
  • Retired Staff
  • Full Member
  • *
  • Posts: 608
  • Mole Snacks: +70/-15
Re:Diagonal Electrophoresis
« Reply #3 on: January 02, 2006, 02:57:18 PM »
Quote
Thank you very much.

You are welcome! Always pleased to help people ;) !
Logged
Pages: [1]   Go Up
« previous next »
Sponsored Links