Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Basic Lab Protocols and Techniques > SDS-PAGE Gel Electrophoresis Forum
Register Search Today's Posts Mark Forums Read

SDS-PAGE Gel Electrophoresis Forum SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


Do you re-use transfer buffer?

Do you re-use transfer buffer? - SDS-PAGE Gel Electrophoresis Forum

Do you re-use transfer buffer? - SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 06-08-2009, 10:08 AM
Anastasia_S's Avatar
Summer Student
Points: 326, Level: 6 Points: 326, Level: 6 Points: 326, Level: 6
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2009
Posts: 70
Thanks: 3
Thanked 12 Times in 12 Posts
Question Do you re-use transfer buffer?



Good time of the day to all,

I had this question after making up yet another bottle of transfer buffer today (for protein transfer from SDS-PAGE gel onto NC membrane).

Can I re-use it? Surely, there are people who tried it, what do you think?

Mine has Tris, glycine, methanol and water, and I usually keep it at 4C. I do a transfer overnight, in a reasonably cold room (probably around 15C), and I cool the tank with running cold water. Do you think this buffer can be used another time(s)?
Reply With Quote
  #2  
Old 06-10-2009, 09:19 AM
Graduate Student
Points: 2,354, Level: 31 Points: 2,354, Level: 31 Points: 2,354, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2009
Posts: 114
Thanks: 19
Thanked 15 Times in 14 Posts
Default Re: Do you re-use transfer buffer?

Never did that before. But I suppose you could try if you want to. To me, a transfer buffer is like a gel electrophoresis buffer. Never reuse.
Reply With Quote
  #3  
Old 06-10-2009, 09:42 AM
Anastasia_S's Avatar
Summer Student
Points: 326, Level: 6 Points: 326, Level: 6 Points: 326, Level: 6
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2009
Posts: 70
Thanks: 3
Thanked 12 Times in 12 Posts
Default Re: Do you re-use transfer buffer?

Thanks, I guess I'll try it later on some not so "precious" samples
Reply With Quote
  #4  
Old 06-11-2009, 02:23 AM
Graduate Student
Points: 2,354, Level: 31 Points: 2,354, Level: 31 Points: 2,354, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2009
Posts: 114
Thanks: 19
Thanked 15 Times in 14 Posts
Default Re: Do you re-use transfer buffer?

Haha! Then do let us know the results after you have tried it!!
Reply With Quote
  #5  
Old 06-11-2009, 03:05 AM
admin's Avatar
Administrator
 
Join Date: Nov 2005
Posts: 1,418
Thanks: 883
Thanked 68 Times in 58 Posts
Default Re: Do you re-use transfer buffer?

yes I have reused transfer buffer quite a bit but make sure you log/dispose of the old buffer every so often.
Reply With Quote
The Following User Says Thank You to admin For This Useful Post:
Anastasia_S (06-14-2009)
  #6  
Old 06-23-2009, 09:19 AM
Anastasia_S's Avatar
Summer Student
Points: 326, Level: 6 Points: 326, Level: 6 Points: 326, Level: 6
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2009
Posts: 70
Thanks: 3
Thanked 12 Times in 12 Posts
Default Re: Do you re-use transfer buffer?

Latest update:
I reused my transfer buffer and it worked fine!
It was stored for 1 week at 4C.
I soaked NC membranes, pads and filter paper in new buffer, just to avoid any undesired protein sticking onto the membrane
Reply With Quote
  #7  
Old 06-23-2009, 09:43 AM
Graduate Student
Points: 2,354, Level: 31 Points: 2,354, Level: 31 Points: 2,354, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2009
Posts: 114
Thanks: 19
Thanked 15 Times in 14 Posts
Default Re: Do you re-use transfer buffer?

great to hear that. though i always use semi dry transfer! haha.
Reply With Quote
  #8  
Old 06-26-2009, 03:43 PM
Pipette Filler
Points: 367, Level: 7 Points: 367, Level: 7 Points: 367, Level: 7
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jun 2009
Posts: 16
Thanks: 1
Thanked 4 Times in 3 Posts
Default Re: Do you re-use transfer buffer?

Quote:
Originally Posted by jiajia1987 View Post
Never did that before. But I suppose you could try if you want to. To me, a transfer buffer is like a gel electrophoresis buffer. Never reuse.
Haha, I am the complete opposite. I always re-use (whether for DNA or protein gels). I usually throw it out after 2-3 uses though. I find it that works perfectly fine if I re-use it fairly soon (within 2 days, stored at 4 C). After a few days, I also tend to toss it. It works fine when I do semi-dry.
Reply With Quote
  #9  
Old 06-29-2009, 01:37 AM
Graduate Student
Points: 2,354, Level: 31 Points: 2,354, Level: 31 Points: 2,354, Level: 31
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2009
Posts: 114
Thanks: 19
Thanked 15 Times in 14 Posts
Default Re: Do you re-use transfer buffer?

Quote:
Originally Posted by jennifer.chang View Post
Haha, I am the complete opposite. I always re-use (whether for DNA or protein gels). I usually throw it out after 2-3 uses though. I find it that works perfectly fine if I re-use it fairly soon (within 2 days, stored at 4 C). After a few days, I also tend to toss it. It works fine when I do semi-dry.

Really? Perhaps I should consider reusing the buffers.

But one thing I do not understand is, when we do semi-dry transfer, the buffer practically gets lost. The transfer buffer is used to soak the membrane and the sponges (that of course can be reused since we can simply soak them by putting them in a small dish with the transfer buffer). However, what happens to transfer buffer that is used to wet the gel? As in, when the sponge, gel and membrane are placed on the transfer, we need to wet it a little more right? Those buffer will be lost since it will be dripped everywhere. You simply reuse the transfer buffer in the dish used to presoak the sponges and membrane, am I right?
Reply With Quote
  #10  
Old 07-06-2009, 03:52 PM
Pipette Filler
Points: 657, Level: 13 Points: 657, Level: 13 Points: 657, Level: 13
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: May 2009
Posts: 27
Thanks: 0
Thanked 7 Times in 6 Posts
Default Re: Do you re-use transfer buffer?

Have you checked if the pH remained the same?
Reply With Quote
Reply

Tags
buffer , nc membrane , protein transfer , reuse , transfer , transfer buffer


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
protein purification magnificientleo Proteomics Forum 11 12-29-2010 09:01 AM
SDS-PAGE Protocol moleculardude Protein Science 14 02-12-2010 11:27 AM
Production of Monoclonal Antibodies - Technique in Mouse Cellbiogal Antibody Forum 2 01-08-2009 10:52 PM
pH of transfer buffer wendyleye Western Blot Forum 0 09-04-2008 01:40 AM
western transfer buffer axexango Protocols and Methods Forum 1 02-05-2005 05:43 PM


All times are GMT. The time now is 06:03 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.16995 seconds with 16 queries