Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protein Science > Western Blot Forum
Register Search Today's Posts Mark Forums Read

Western Blot Forum Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


Western Blot Stoping PointS!! ADD YOURS

Western Blot Stoping PointS!! ADD YOURS - Western Blot Forum

Western Blot Stoping PointS!! ADD YOURS - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 05-02-2011, 11:28 PM
Pipette Filler
Points: 358, Level: 7 Points: 358, Level: 7 Points: 358, Level: 7
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2011
Location: SF
Posts: 19
Thanks: 10
Thanked 5 Times in 5 Posts
Default Western Blot Stoping PointS!! ADD YOURS



We've all had that wanting to stop our western, the two day experiment but afraid to loss what little progress we've made.....so please put your thoughts here tried and true places to stop a western without effecting results!!
Reply With Quote
  #2  
Old 05-03-2011, 05:30 AM
Volunteer
Points: 960, Level: 17 Points: 960, Level: 17 Points: 960, Level: 17
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Oct 2010
Location: spain
Posts: 58
Thanks: 1
Thanked 27 Times in 25 Posts
Default Re: Western Blot Stoping PointS!! ADD YOURS

Hola, I think that is better run the gel and transfer it at the same day. but after it you can stop in any of steps.Blocking, you can let the WB blocking ON at 4║, in this case more clean will be the final image. First ab.: same you can let it ON at 4║, once the maximum bounding is reach the signal doesn┤t increase , and second ab similar but have the risk of lost of activity of perosidase, but at 4║ it┤s improbable and if you are washing and have to go , the same you can extend one wash ON or a weekend if you want. Wait for more opinions . Buena suerte
Reply With Quote
The Following 2 Users Say Thank You to protolder For This Useful Post:
admin (05-04-2011), mai-mb (05-04-2011)
  #3  
Old 05-03-2011, 06:17 PM
danfive's Avatar
Nobel Laureate
Points: 8,612, Level: 64 Points: 8,612, Level: 64 Points: 8,612, Level: 64
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2007
Location: Houston TX
Posts: 969
Thanks: 12
Thanked 184 Times in 154 Posts
Default Re: Western Blot Stoping PointS!! ADD YOURS

I like to dry my blot paper after the transfer, air dry at RT on paper towel, protein side up.
I'll store same way the whole weekend if needed.
Reply With Quote
The Following 2 Users Say Thank You to danfive For This Useful Post:
admin (05-04-2011), mai-mb (05-04-2011)
  #4  
Old 05-04-2011, 05:45 PM
Pipette Filler
Points: 358, Level: 7 Points: 358, Level: 7 Points: 358, Level: 7
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2011
Location: SF
Posts: 19
Thanks: 10
Thanked 5 Times in 5 Posts
Default Re: Western Blot Stoping PointS!! ADD YOURS

Awesome thanks you guys your expertise is highly appreciated!! Ive tried both the o/n primary incubation yesterday with good results today. And will try the dry membrane storage probably this weekend.
Reply With Quote
Reply

Tags
add , blot , points , stoping , western


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
Western Blot Background Troubleshooting molecule2005 Western Blot Forum 18 10-07-2010 11:49 AM
Can we use dot blot inplace of Western blot? peterish Western Blot Forum 3 06-16-2010 07:59 PM
Western blot admin Article Discussion 2 04-23-2010 07:31 PM
dot blot and western blot Liang Western Blot Forum 1 08-07-2009 03:26 PM
Qi: Standard Model Extension or Fifth Force? cinquirer Physics Forum 88 11-21-2003 10:45 PM


All times are GMT. The time now is 04:22 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.13013 seconds with 16 queries