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

Immunohistochemistry Forum Immunohistochemistry Forum. Post and discuss questions on immunohistochemistry methods including paraffin, frozen, or free floating tissue sections.


Reusing Primary Antibody in Immunohistochemistry

Reusing Primary Antibody in Immunohistochemistry - Immunohistochemistry Forum

Reusing Primary Antibody in Immunohistochemistry - Immunohistochemistry Forum. Post and discuss questions on immunohistochemistry methods including paraffin, frozen, or free floating tissue sections.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 07-11-2008, 10:50 PM
Pipette Filler
Points: 2,218, Level: 30 Points: 2,218, Level: 30 Points: 2,218, Level: 30
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2006
Posts: 10
Thanks: 0
Thanked 0 Times in 0 Posts
Default Reusing Primary Antibody in Immunohistochemistry



Hello, can we also reuse primary antibodies in immunohistochemistry IHC? I know that we can reuse primaries after westerns, but i would like to reuse the IHC antibodies to save money. Does anyone have experience in this?
thanks
Reply With Quote
  #2  
Old 04-23-2009, 10:28 PM
Pipette Filler
Points: 48, Level: 1 Points: 48, Level: 1 Points: 48, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Apr 2009
Posts: 9
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Reusing Primary Antibody in Immunohistochemistry

Normally we draw from the concentrated antibody vial only what we need for the procedure at hand. We prepare the working dilution from this draw, thus leaving the concentrated antibody in the original vial for further use. We do not re-use the dliuted antibody that has incubated with the test tissue.
Reply With Quote
  #3  
Old 08-01-2009, 09:36 AM
Pipette Filler
Points: 44, Level: 1 Points: 44, Level: 1 Points: 44, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Aug 2009
Posts: 3
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Reusing Primary Antibody in Immunohistochemistry

Quote:
Originally Posted by orestis View Post
Hello, can we also reuse primary antibodies in immunohistochemistry IHC? I know that we can reuse primaries after westerns, but i would like to reuse the IHC antibodies to save money. Does anyone have experience in this?
thanks
Hi radam. Maybe you already solved the problem. We prepare dilutions for about 20-30 samples, with antibodies we use everyday, and they work for at least 1 month if you keep it in the fridge, and avoiding high temperatures, sunlight and a long time exposure to air oxidation. We also alicuotate dilutions for some special antibodies and keep them at -20 degrees.
Reply With Quote
  #4  
Old 11-12-2009, 01:06 AM
Pipette Filler
Points: 29, Level: 1 Points: 29, Level: 1 Points: 29, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Nov 2009
Posts: 8
Thanks: 1
Thanked 0 Times in 0 Posts
Default Re: Reusing Primary Antibody in Immunohistochemistry

yes i agree with radam and hunydd because after incubating the tissue sections with primary antibody, the concentration of antibody left in the solution will be different, i.e lower, and it may or may not give you consistent staining quality!
Reply With Quote
  #5  
Old 03-12-2013, 09:33 PM
haisond's Avatar
Pipette Filler
Points: 80, Level: 1 Points: 80, Level: 1 Points: 80, Level: 1
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2013
Location: los angeles, CA
Posts: 14
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Reusing Primary Antibody in Immunohistochemistry

Airie is correct, with each reuse, the antibody concentration decreases due losses by binding of antibodies to target antigens. If you still get good results with a reuse, that means that your original primary ab dilution was very concentrated. With primary antibodies there are no ways to stretch the buck here with sacrificing quality.
Reply With Quote
Reply

Tags
antibody , immunohistochemistry , primary , reusing


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
can I use mouse antibody to do immunohistochemistry in mouse brain sections? llzz Antibody Forum 1 03-12-2013 09:02 PM
Production of Monoclonal Antibodies - Technique in Mouse Cellbiogal Antibody Forum 2 01-08-2009 10:52 PM
primary vs secondary antibody public_enemee Western Blot Forum 1 10-31-2008 06:01 AM
Western background staining with Primary antibody (Rabbit) Sharon Waldrop Protocols and Methods Forum 2 08-19-2008 01:44 PM
Primary antibody species minusach Antibody Forum 1 06-28-2007 01:42 PM


All times are GMT. The time now is 10:48 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 0.63549 seconds with 16 queries